CONTENT

S.NO TITLE PAGE.NO

1 INTRODUCTION 1

2 MATERIALS AND METHODS 42

3 RESULT AND DISCUSSION 66

4 SUMMARY 75

5 REFERENCES 78
Diabetes mellitus:

In most instances, Diabetes mellitus results from diminished secretion

of insulin by the β-cells of the islets of langerhans. Hereby usually plays a

major role in determine in whom diabetes will develop and in whom it will

not. Sometimes it does this by increasing the susceptibility of the β-cells to

destruction by viruses or by favoring the development of autoimmune

antibodies against the β-cells, thus also leading to their destruction. In order

instances, there appears to be a simple hereditary tendency for β-cell

degeneration.

Obesity also plays a role in the development of clinical diabetes. One

reason is that obesity decreases the number of insulin receptors in the insulin

target cells through the body, thus making the amount of insulin that is

available less effective in promoting its usual metabolic effects.(Guyton, C

John E.Hall, et.al.1998)

Type I DM: Insulin dependent diabetes mellitus (IDDM)

 Type I diabetes usually develops before the age of 40 and hence is

called juvenile diabetes. Patients with this disease are not obese, and they

have a high incidence of ketosis and acidosis, various anti B cell antibodies

are present in plasma, but the current thinking is that type I diabetes is

primarily a T-lymphocyte mediated disease. There is a definitive genetic

susceptibility as well; if one identical twin develops the disease; There is a

one –in – three chances that the other twin will do so also. In other words,

the concordance rate is about 33%. The main genetic abnormality is in the

major histocompatibility complex on chromosome 6, making individuals

with certain types of histocompatibility antigen much more prone to develop

the disease. Other genes are also involved.

Immunosuppression with drugs such as cyclosporine ameliorate type I

diabetes if given early in the disease before all B cells are lost. Attempts

have been made to treat type I diabetes by transplanting pancreatic tissues

or isolated islet cells, but results to date have been poor, largely because B

cells are easily damaged made it is difficult to transplant enough of them to

normalize glucose responses. (William F. Ganong, M.D; 2005).

Type II Non insulin dependent diabetes mellitus (NIDDM)

NIDDM is the most common form of Diabetes. Typically, this

disease is characterized by an underproduction iof insulin or insulin

insensitivity at target tissues. Genetic factors may play a greater role in this

disease than in IDDM. Concordance rates for NIDDM in identical twins are

100%. Currently, a genetic marker has not been discovered for NIDDM. The

etiology of this disease is unknown, although some observed correlations

may eventually assist in determining its causes, Environmental factors, such

as diet may play an important role in the development of NIDDM.

Obesity, especially of the abdominal viscera, is common in

individuals with NIDDM. Changes in the normal substrate concentrations

delivered to the liver, adipose tissue, and skeletal muscle are thought to

affect the normal functioning of the insulin receptor (Haring HU. 1991).

This in turn may decrease the number of glucose transports that are

delivered to the cell surface, thereby decreasing the delivery of glucose into

the cell. Although this model oversimplifies the actual molecular

mechanisms, if is known that restriction of caloric intake and weight less are

often sufficient to control NIDDM.

Otherwise, the patient is usually treated with sulfonylureas or other

oral hypoglycemic drugs. These drugs appear to facilitate the release of

insulin from beta cell and may also increase the target tissues, sensitivity to

insulin. Interestingly, NIDDM may be, in part, a disease caused by an

imbalance between insulin and glucagons concentrations. It has been

observed in true NIDDM individuals that the numbers of beta cells do not

decrease: however, there is a significant increase in the number of alpha

cells in the islets.(Unger RH: Foster DW, et. Al 1992).

Gestational diabetes

Gestational diabetes refers to diabetes that occurs temporarily during

pregnancy. One study estimates that 39% of women with gestational

diabetes manifest type II diabetes mellitus 20 years after delivery. Screening

of pregnant women for gestational diabetes, to prevent perinatal

complications associated with maternal hyperglycemia, has become a

widespread accepted practice. (O’ Sullivan JB, Worshop, et al 1984).

Insulin

Insulin is synthesized in the endocrine pancreas by the β-cells of the

islets of Langerhans as a high molecular- weight precursor called

preproinsulin (Chan SJ. Keimp, stenor DF, e.al 1976) cleavage at the link

marked by the arrow labeled I results in the formation of proinsulin (9000D).

Proinsulin has only 5% of the activity of insulin. The proinsulin molecule

consists of A and B chain if insulin connected by disulfide bonds and by a

connective peptide called c-peptide. During processing the c-peptide

(3000D) is removed from the molecules by cleavage at the links marked by

arrows 2 and 3. The resulting insulin molecule (6000D) consists of chain A

and B connected by two disulfide bonds. This entire process occurs within

the β-cell. The initial synthesis of preproinsulin occurs at the Golgi

apparatus. The molecule is packaged in a vesicle Called β granule.

Cleavage first proinsulin and next to insulin occurs within are released into

the circulation when the granule is dissolved at the plasma membrane of the

β-cell after neural, dietary, or hormonal stimuli. Only small quantities of

proinsulin are found in the circulation.

Insulin synthesis and secretion

Insulin consists of two peptide chains linked by two disulfide

bonds. The α chain contains 2 amino acids and the β chain 30 amino acids.

The molecular weight of insulin monomer is 5500 Da. The precursor of

insulin within the β cells of the islet of Langerhans is the single chain

preproinsulin. During insulin synthesis a 2- amino acids signal sequence is

first cleaved from preproinsulin by a peptidase, yielding proinsulin.

Proinsulin and c-peptide

 Proinsulin consists of the insulin sequence interspersed by a

connecting peptide (c-peptide). At the final stage of insulin synthesis,

proinsulin is split into insulin and C-peptide, both of which are then released

from the cell. C-peptide is released in an amount equimolar to insulin. This

is exploited in the clinical laboratories to assess β cell function in patients

treated with exogenous (therapeutically injected) insulin. In these patients,

endogenous insulin cannot be measured directly, because the exogenous

insulin would interference in the assay. In such circumstances, C-peptide

measurement provides an assessment of B-cell function.(Baynes,

Dominiczalc, et al 1999).

Insulin receptor

Insulin binds to a receptor on the plasma membrane of target cells, and

when this insulin-receptor complex form, glucose is allowed to enter into the
target cell where it is used as an energy source or is converted into glycogen
for energy storage. The insulin receptor is a protein consisting of two copies
of two different peptide units, alpha and beta. One insulin molecule is
required to bind to each alpha subunit, and after this occurs, the ß subunits
then transmit a signal that causes the cytoplasmic end of the receptor protein
to change shape. This change causes a cytoplasmic protein kinase active site
to be exposed which causes the phosphorylation of an insulin receptor
substrate which initiates other reactions that lead to the uptake of glucose by
the cell (Purves et al, 2000). This process is shown in Figure.

Figure 3: Insulin-receptor complexes on the cell surface cause chemical

responses to occur within the cell. This figure was reproduced pending
permission from the authors of Life, 6th Ed (Purves et al, 2001).

When the concentration of glucose in the blood is high, the pancreas

releases insulin into the bloodstream. The insulin travels throughout the
bloodstream until it finds its receptors on the surface of cells. After two
insulin molecules bind to the insulin-receptor complex, a series of complex
reactions occurs which ultimately leads to the uptake of glucose by cells.
These cells are then able to use glucose as an energy source. For example,
fat cells use glucose to produce fat and liver cells use glucose to produce
glycogen and fat to be stored for later energy use. When the concentration of
glucose in the bloodstream is low, most cells in the body use glycogen and
fat as their energy source instead of glucose (Purves et al, 2001).

Receptor site defects

Insulin receptor protein

Insulin binds reversibly to sites on cell membrane. Insulin binding sites

(called receptor sites) are found only on certain cell types (liver cells,

monocytes, adipocytes, and muscle). The insulin receptor site is

composed of two glycoprotein molecules. (Kasuga M.Van obberghan, et al

1981). One subunit is a tyrosine – specific protein kinase (Roth. RA. Cassell

et al 1983). Insulin binding to the receptor site triggers a chain of events

resulting in an increase of cell membrane permeability to glucose and amino

acids, alteration of enzyme activities, and promotion of protein biosynthesis.

It is low relative to both the blood glucose and insulin levels. Insulin

resistance in type II diabetes is directly related to a decreased numbers of

insulin-receptors.(Moller DE, Filer JS, et al 1991). Diabetes caused by

decreased numbers of insulin - receptors sites is called type A diabetes.

Type A diabetes occurs obese person. Obese individuals show a significant

increase in the numbers of insulin-binding sites and a decrease in symptoms

of diabetes when placed on a low-carbohydrates diet. Type A diabetes in

obese persons may derived directly from a high carbohydrate diet rather than

obesity per se.

Antibodies to receptor
The presence of circulating antibodies to the insulin receptor has also
been reported (R). Type II diabetes caused by such an antibodies to insulin
receptor sites is called type B diabetes. Type B diabetes usually has
symptoms of autoimmune disorders such as antinuclear antibodies
arthralgia, and enlargement of the parotid gland. Type B Diabetes has a
lower incidence than type A

Impaired glucose transport

Glucose transport is reduced in both type I and type II diabetes because
of significantly reduced levels of the high Km glucose transport protein
GluT-2 (Unger RH 1991).

Insulin resistance
Insulin resistance is a condition in which the body produces insulin but
does not use it properly. When people are insulin resistant, their muscle, fat,
and liver cells do not respond properly to insulin. As a result, their bodies
need more insulin to help glucose enter cells. The pancreas tries to keep up
with this increased demand for insulin by producing more. Eventually, the
pancreas fails to keep up with the body’s need for insulin. Excess glucose
builds up in the bloodstream, setting the stage for diabetes. Many people
with insulin resistance have high levels of both glucose and insulin
circulating in their blood at the same time.
Etiology of diabetes mellitus
Environmental, genetic and autoimmune factors involved in the
etiology of diabetes (Michael, R.C., 1992).
1. Induction of diabetes mellitus by environmental agents

Environmental agents may have deleterious effect, on β-cells

number of chemical are known to have relatively specific cytotoxic effect
on β-cells including alloxan, stroptozotocin and rodenticide racor (Yoon,
J.W., Kim, C.J., Pake , Y) encephalomgo- cariditis, Virus and causative β-
virus and can induce diabetes in animal by infecting Pancreatic β -cells
(Taylor R, AgiusL, et al.)Tropism and diabetogenic potential in animal and
the patients with congenital rubella syndrome, particularly those with HLA-
DR3 (Pyorula K and Laakso M, e.al )

2. Induction of diabetes mellitus by genetic factors

Both type I and type II diabetes are at least partly inherited Type I
diabetes appears to be triggered by some mainly viral infections or less
likely stress-related or environmental factors. There is a genetic element in
individual susceptibility to some of these triggers which has been traced to
particular HLA genotypes who have inherited the susceptibility type I
diabetes mellitus seems to require an environmental trigger. A small
proportion of people with type I diabetes carry a mutated gene that cause
maturity onset diabetes of the young (Mody)
 There is a stronger inheritance pattern for type II diabetes. Those with
first degree relatives with type II have a much higher risk of developing
type II concordance among monozygotic twins is close to 100% and 25%
of these with the disease have a family history of diabetes. This is found
approximately 85% of the patients diagnosed. So some experts believe that
inheriting a tendency towards obesity may also contribute.

3. Induction of diabetes mellitus by autoimmunity

Autoimmune reactions against one or more components of pancreatic

β-cells result in the destruction of the beta cells (Sttahn R.M, Gohdes D. et
al 1993). These observations suggest that destructive process leading to
IDDM is probably immune mediated. The viral infection causing human
diabetes is evident from the studies of patients with congenital rubella
syndrome (Farell, M.A., QuigginsP.A, et al 1993). Children infected
congenitically may show rubella virus in the pancreas with insulin and β-
cells destruction (Kleinman, J, C., Donahere, R.B). More convincing
evidence for viral involvement is derived from the study of young boy who
died from overwhelming viral infection and diabetic ketoacidosis. (Karamj,
H., Lewitt, P.A. 1993).

Clinical Features of diabetes mellitus

1. Glucosuria
Loss of glucose in urine is known as glucosuria normally glucose dose
not appear in urine. When glucose level rises above 180mg/dl in blood
glucose appears in urine. It is the renal threshold level for glucose.
2. Osmotic diuresis
Diuresis due to osmotic effects is called osmotic dieresis. The excess
glucose in the renal tubules develops osmotic effect. The osmotic effect
decreases the reabsorption of water from renal tubules resulting in dieresis.
It leads to polyuria and polydipsia.

3. Polyuria
Excess urine formation with increase in frequency of vooling urine is
called polyuria. It is due to the osmatic diuresis caused by increase in blood
sugar level.

4. Polydipsia
The increase in water intake is called polydipsia. The excess loss of
water decreases water content and increases salt content in the body. This
stimulates the thirst center in hypothalamus. Thirst center in turn increases
the intake of water.

5. Asthenia
The loss of strength is called asthenia. The body becomes very weak.
There is loss of energy. Asthenia is because of lack of insulin, which causes
decrease in protein syntheses and increase in protein breakdown. Protein
depletion also occurs due to the utilization of proteins for energy in the
absence of glucose utilization.

6. Acidosis
 During insulin deficiency glucose cannot be utilization by the peripheral
tissues for energy. So a large amount of fat is broken down to release
energy. It causes the formation of excess ketoacidosis leading to acidosis.
One more reason for acidosis is that, the ketoacidosis are
excreted in combination with sodium irons through urine (ketonuria).
Sodium in exchanged for hydrogen ions which diffuse from the renal tubules
into ECF adding to acidosis.

7. Polyphagia
Polyphagia means the intake of excess food. It is very common in
diabetes mellitus.
8. Acetone breathing
In cases of severe ketoacidosis, acetone is expired in the expiratory air,
giving the characteristic atone or fruity breath odor. It is a life-threatening
condition of severe diabetes.

9. Kussmaul breathing
Severe acidosis increases the rate and depth of respiration which is
known as Kussmaul breathing.

10. Circulating shock

The osmotic diuresis leads to dehydration, which causes circulatory
shock. It occurs only in severe diabetes.

11. Coma
Due to Kussmaul breathing, large amount of carbon dioxide is lost
during expiration. It leads to drastic reduction in the concentration of
bicarbonate ions causing severe acidosis and coma. It occurs in severe cases
of diabetes mellitus.
Increases in blood sugar level develop hyperosomolarity of plasma
which also leads to coma. It is called hyperosmolar coma.(Medical
physiology.,sembulingam.,fourth edition 2006)

Diagnosis of DM
The acute state of IDDM is readily diagnosed from the history,
symptoms (hunger, thirst, frequent urination, weight loss) and the
following abnormal laboratory tests: plasma glucose (>200mg /dL, 11.1m
mol/L), glucosuria (usually 4+or 2g/day), and presence of ketone bodies in
urine and serum. The asymptomatic patient presents a diagnostic problem,
but the tests described in the following paragraphs are helpful.

Fasting plasma Glucose

The diabetes Data group (National Diabetes data group 1979)
proposed that a fasting (overnight) plasma glucose concentration exceeding
140mg/dL (7, 8 mol/L) on 2 separate occasions should be accepted as a
criterion for diabetes. Others have proposed the higher limit of 150mg/dL.
Milder cases of hyperglycemia require some type of carbohydrate loading
test for detection. (Ito. C., Mito K. et al 1983).

Postprandial plasma glucose

The simplest leading test is the measurement of plasma glucose

concentration 2 hour after the patient consumes a meal containing
approximately 100g of carbohydrate mixed with other foodstuffs. A
 plasma glucose value exceeding 200mg/dL is indicative of diabetes.
Whereas a value below 120mg/dL is considered normal. Concentrations
between 120 and 200mg/dL are equivocal and further study.(Alex Kaplan,
Rhona jacle et al 1995).

O’Sullivan test

The O’ Sullivan test is frequently used to detect gestational diabetes.

A 50g load of glucose is given to a fasting patient. Blood is drawn at 1
hour. Gestational diabetes is suggested by plasma glucose levels above
1500 mg/L above 1300 mg/L for whole blood.

The oral glucose tolerance test

The oral glucose tolerance test (OGTT) is a standard for making the
diagnosis of type II diabetes. It is still commonly used for diagnosing
gestational diabetes and in conditions of pre-diabetes, such as polycystic
ovary syndrome with an oral glucose tolerance test.

Glycosylated haemoglobin or HbAIC

While nor used for diagnosis, is an elevated level of glucose bound

to haemoglobin (termed glycosylated haemoglobin or HbAIC) of 60% of
higher considered abnormal by most labs: HbAIC is primarily a treatment
tracking test reflecting average blood glucose levels over the proceeding 90
days. However, some physician may order this test at the time of diagnosis
to track changes over times. The current recommended goal for HbIAC in
patients with diabetes is less than 70% as defined as “goal glycemic control
“although some guidelines are striker (<6.5%). People with diabetes that
have HbAIC levels with this goal have a significantly lower incidence of
complications from diabetes, including retinopathy and diabetic nephropathy
(Genuth, S.2006).

COMPLICATION OF DIABETES MELITUS:

RETINOPATHY
Opaque areas in the lens of the eye are called cataracts. Cataract
formation is the principle retinopathy of diabetes. Retinopathy is also caused
by proliferation of small blood vessels in the lens. (Wrolewski A .S, Warram
JH, et al 1987).

NEUROPATHY:
Neuropathy is the most common complication of Diabetes mellitus.
It is apparent in about 25% of Diabetes and is recognized by a variety of
symptoms that include pain, numbness, tingling or burning sensation in
extremities, dizziness and double vision. These symptoms are caused by
decreased motor and sensory nerve deduction velocities cased by axonal
degeneration and demyelization secondary manifestations of neuropathy
include cardiac failure excessive sweating and male impotence (Editorial
Lancet. 1993)

ANGIOPATHY
Angiopathy refers to damage lining (basement membranes) of blood
vessels. Angiopathy increases the risk of coronary heart disease and stroked
and can lead to retinopathy and nephropathy.
NEPHROPATHY
 Nephropathy refers to damage to the glomerulus’s (filtering
apparatus of the nephron) and capillaries associated with the glomerulus’s
capillaries associated with the glomerulus’s capillary damage is cased by
antipathy. The result is reduction in the filleting capability of the kidneys
proteinuria is often the fist sign of diabetic nephropathy. Approximately 25-
30%of individual treated for end-stage renal failure are diabetics.

INFECTION
Diabetics are highly susceptible to infection, ulceration, and gangrene
(especially in the extremities) skin disorders are also more common in
diabetics than in non-diabetics,

HYPERLIPIDEMIC AND ATHEROSCLEROSIS

High triglyceride and cholesterol levels are often associated with

type II diabetes (Bradky RF, Krall Lp, et al 1971). Increased levels of very
–low density lipoproteins (VLDL) have been reported for type II diabetics
(Goldberg RB. 1981). High density lipoproteins (HDL) have been reported
to be significantly lower in diabetes than in non -diabetics (Lopes-virells
MFL, stone PG, et al 1977). These results are consistent with the higher
incidence of coronary heart disease in diabetics and a poor survival rate for
diabetics with myocardial infraction (Smith Jw, Marcus FI, et al.)

Metabolic changes in diabetes mellitus

Hyperglycemia

Occurs as a result of,

 • Decreased and impaired transport and uptake of glucose into muscles
and adipose tissues.
• Repression if key glycolytic enzymes like glucokinase,
phosphofructokinase and pyruvate kinase.

• Depression of key gluconeogenic enzymes like gflucose-6-

phosphatase, fructose 1,6 bisphosphatase, pyruvate carboxylase,
phosphoenol pyruvate carboxy kinade occur promoting
gluconeogenesis in liver.

Amino acid level

Transport and uptake of amino acid in peripheral tissues is also
depressed causing an elevated circulating level of amino acids. Amino
acids. Amino acids breakdown in liver results in increased production of
urea.

Protein synthesis
Protein is decreased in all tissues due to
• Depressed production of ATP
• Absolute or relative deficiency of insulin

Fat metabolism
Stored lipids are hydrolyzed by increased lipolysis liberating
free fattyacids. Increased free fatty acid interferes at several steps of
carbohydrate and phosphorylation in muscles, further contributing to
hyperglycemia. Excessive production of ketone bodies increases the
concentration of ketone bodies in blood and excretion of ketone bodies in
urine and leads to acidosis.

Effect on glycogen synthesis

Glycogen synthesis is depressed as a result of
• Decreased glycogen synthase activity due to deficiency of insulin
• By activation of phosphorylase
• By producing glycogenolyses through the action of epinephrine or
glucagons

CARBOHYDRASE INHIBITORS

The group of agents known as carbohdrase inhibitors includes

acarbose, which is available for therapy in Europe, and miglitol, which is
still, is clinical trials. (Bresserl R, Johnson D, et al. 1992) The mechanism
of action of both drugs is inhibition of the-α glycosidase in the intestinal
brush border, leading to a delay in carbohydrate absorption. Clinical studies
with these agents have demonstrated a reduction is postprandial glucose
elevarions in both insulin dependent diabetes mellitus and NIDDM.
(Vierhapper H, Bratusch marrain A. et al 1978) Doses of the carbohydrase
inhibitors must be titrated to balance the decreased glycemic response
against malabsorption and other gastrointestinal side effects. For both
agents, the therapeutic dose is between 50 and 100 mg: higher doses lead to
abdominal distention, flatulence, malabsortion and diarrhea. Both these
agents have also been used in combination with sulfonylurea. (Johnston PS
Coniff FR et al 1994). The use of these agents has generally been associated
with modest improvement in gylcosylated haemoglobin measurements.

OTHER STRATEGIES FOR DRUG THERAPHY

Several other strategies for treating NIDDM are under investigation.

(Calrk BF, Duncan LPJ. et al 1979.) Fatty acid oxidation inhibitors should
prevent the actions of fatty acids in stimulating hepatic gluconeogenesis and
in attenuating glucose disposal in muscle, as well as diminished rates of
ketone, cholesterol, and triglyceride synthesis. (Foley JE. et al 1992). The
potential usefulness of inhibitors of gluconeogenesis in also being studied.
Another strategy involves the synthesis of glucagons analogous that can
antagonize the action of glucagons. (Bregman MD, Trivedi D, et al 1980.)
Finally, peptides with insulinotropic effects, such as glucagons- like
peptides, are also under investigation. The ideal therapy for NIDDM will
probably be achieved when the fundamental pathogenesis of the disease is
better understood.

TREATMENT OF NONKETOTIC HYPEROSMOLAR COMA

Fluid repletion is the most important aspect of treatment. The deficit,

which may reach 10L or more, exceeds that of diabetic ketoacidosis. (Arief
AI, Carroll HJ, et al 1972). The first 2 or 3L should be given rapidly, even
in elderly patients with uncertain cardiac function. Careful monitoring of
the central venous pressure permits rapid repletion of volume without a risk
of overexpansion. The initial serum sodium level may be high, normal, or
low, depending on the relative losses of sodium and water in the urine
caused by a shift of water out of cells secondary to hyperglycemia.
Treatment should begin with normal saline at a rate that will replenish
at least half of the estimated fluid deficit within 6h, after which 0.45%
saline can be given to complete volume replacement. Re-expansion of the
extra cellular fluid volume reduces of level of glucagons, catecholamines,
and the other hormones of stress and re-establishes glucose excretion in
renal function is intact. (Foster DW et al 1983.). Although fluids reduce
hyperglycemia, insulin should also be given. A low-dose schedule
consisting of a 10-v bolus and 5 to 10v/h thereafter is appropriate.

Insulin can precipitate vascular collapse if it causes a major shift of

extra cellular fluid into cells or causes a capillary leak syndrome. Insulin
therapy should always be preceded or accompanied by appropriate
administration of fluids. Because of the high rate of infection, particularly
with gram –negative organism, antibiotics should be given empirically to
any patient with fever pending the outcome of blood, urine, or sputum
(transtraheal aspirate) cultures. Although mortality rats are generally high in
hyperosmolar coma (>50%) lower rates (14%) are reported by some.
(Carrol, et al) Fatalities are highest in older patients.

Physiological Treatment
Exercise and physical activity.
In general, the more active you are, the lower your blood sugar.
Physical activity causes sugar to be transported to your cells, where it's used
for energy, thereby lowering the levels in your blood. Aerobic exercises such
as brisk walking, jogging or biking are especially good. But gardening,
housework and even just being on your feet all day also can lower your
blood sugar.

• Medications. Insulin and oral diabetes medications deliberately work

to lower your blood sugar. But medications you take for other
conditions may affect glucose levels. Corticosteroids, in particular,
may raise blood sugar levels. Medications such as thiazides, used to
control high blood pressure, and niacin, used for high cholesterol, also
may increase blood sugar. If you need to take certain high blood
pressure medications, your doctor will likely make changes in your
diabetes treatment.
• Illness. The physical stress of a cold or other illness causes your body
to produce hormones that raise your blood sugar level. The additional
sugar helps promote healing. But if you have diabetes, this can be a
problem. In addition, a fever increases your metabolism and how
quickly sugar is utilized, which can alter the amount of insulin you
need. For these reasons, be sure to monitor your glucose levels
frequently when you're sick.
• Alcohol. Even a small amount of alcohol? About 2 ounces? Can cause
your sugar levels to fall too low. But sometimes alcohol can cause
sugar levels to rise. If you choose to drink, do so only in moderation.
And monitor your blood sugar before and after consuming alcohol to
see how it affects you. Also, keep in mind that alcohol counts as
• carbohydrate calories in your diet.
• Fluctuations in hormone levels. The female hormone estrogen
typically makes cells more responsive to insulin, and progesterone
 makes cells more resistant. Although these two hormones fluctuate
throughout the menstrual cycle, the majority of women don't notice a
corresponding change in blood sugar levels. Those who do are more
likely to experience changes in blood sugar during the third week of
their menstrual cycle, when estrogen and progesterone levels are
highest.

Hormone levels also fluctuate during perimenopause? The time before

menopause. How this affects blood sugar varies, but most women can
control any symptoms with additional exercise and changes in their diet. If
your symptoms are more severe, your doctor may recommend oral
contraceptives or hormone replacement therapy (HRT). After menopause,
many women with diabetes require about 20 percent less medication because
their cells are more sensitive to insulin.

A healthy diet

Contrary to popular myth, there's no "diabetes diet." Furthermore,

having diabetes doesn't mean you have to eat only bland, boring foods.
Instead, it means you'll eat more fruits, vegetables and whole grains? foods
that are high in nutrition and low in fat and calories ? And fewer animal
products and sweets. Actually, it's the same eating plan all Americans should
follow.

Yet understanding what and how much to eat can be a challenging task.
Fortunately, a registered dietitian can help you put together a meal plan that
fits your health goals, food preferences and lifestyle. Once you've decided
on a meal plan, keep in mind that consistency is extremely important. To
keep your blood sugar at a consistent level, try to eat the same amount of
food with the same proportion of carbohydrates, proteins and fats at the
same time every day.

But even with all the information you need and the best intentions,
sticking to your diet can be one of the most challenging parts of living with
diabetes. The key is to find ways to stay motivated. Don't let others
undermine your determination to eat in the healthiest way possible. You
have to believe that what you're doing matters? And that you're worth it.

Healthy weight

Being overweight is the greatest risk factor for type 2 diabetes. That's
because fat makes your cells more resistant to insulin. But when you lose
weight, the process reverses and your cells become more receptive to
insulin. For some people with type 2 diabetes, weight loss is all that's needed
to restore blood sugar to normal. Furthermore, a modest weight loss of 10 to
20 pounds is often enough.

Yet losing even 10 pounds can be a challenge for most people.

Fortunately, you don't have to do it alone. A registered dietitian can help you
develop a weight-loss plan that takes into account your current weight,
activity level, age and overall health. Ultimately, however, the motivation
has to come from you.

Oral Diabetes Medications

Usually, people with Type 1 diabetes don't use oral medications. These
medications work best in people with Type 2 diabetes who have had high
blood sugar for less than ten years and who have normal weight or obesity.
It's not uncommon for oral medication to control blood sugar well for years
and then stop working. Some people who begin treatment with oral
medications eventually need to take insulin.

Sulfonylureas

Until 1994, sulfonylureas were the only oral medications for

diabetes available in the US. These medications act to force your pancreas to
make more insulin, which then lowers your blood sugar.
For this medication to work, your pancreas has to make some
insulin. If your pancreas makes no insulin at all, you aren't a good candidate
for this class of drugs. Also, if you have an allergy to sulfa drugs you should
probably avoid sulfonylureas.

Side Effects

• low blood sugar

• an upset stomach
• skin rash or itching
• weight gain

Biguanides
The generic (non-brand) name of this drug is metformin. It helps lower
blood sugar by making sure your liver doesn't make too much sugar.
Metformin also lowers the amount of insulin in your body. You may lose a
few pounds when you start to take metformin. This weight loss can help you
control your blood glucose. Metformin can also improve blood fat and
cholesterol levels, which are often high if you have Type 2 diabetes.
Side Effects

• Lactic acidosis is rare,

• Diarrhea, nausea

Alpha-glucosidase inhibitors

There are now two alpha-glucosidase inhibitors, acarbose and

miglitol. Both medications block the enzymes that digest the starches you
eat. This action causes a slower and lower rise of blood sugar through the
day, but mainly right after meals.

Side Effects
Taking this medication may cause stomach problems such as gas,
bloating and diarrhea that most often go away after you take the medication
for awhile.

Thiazolidinediones

The generic names for these medications are pioglitazone and

troglitazone. The medication works by helping make your cells more
sensitive to insulin. The insulin can then move glucose from your blood into
your cells for energy.

Side Effects
 Edema, weight gain, decreased hematocrit and hemoglobin,
elevated alanine aminotransferase activity, hypoglycemia- during
combination therapy with sulfonylureas or insulin. Swelling in the legs or
ankles.

Meglitinides

This is a new type of diabetes medication. Repaglinide is a generic (non-

brand) name for one of the meglitinides. This medication helps your
pancreas make more insulin right after meals, which lowers blood sugar.
Your doctor might prescribe repaglinide by itself or with metformin if one
medication alone doesn't control your blood sugar a good thing about
repaglinide is that it works fast and your body uses it quickly. This fast
action means you can vary the times you eat and the number of meals you
eatmoreeasilythan you can with other diabetes medications. These work like
short acting Sulfonylureas.

Side Effects

• weight gain
• low blood sugar

Transplantation
In recent years, researchers have focused increasing attention on
transplantation for people with type 1 diabetes. Current procedures include:

• Pancreas transplantation. Pancreas transplants have been performed

since the late 1960s. Most are done in conjunction with or after a
kidney transplant. Kidney failure is one of the most common
 complications of diabetes, and receiving a new pancreas when you
receive a new kidney may actually improve kidney survival.
Furthermore, after a successful pancreas transplant, many people with
diabetes no longer need to use insulin. Unfortunately, pancreas
transplants aren't always successful. Your body may reject the new
organ days or even years after the transplant, which means you'll need
to take immunosuppressive drugs the rest of your life. These drugs are
costly and can have serious side effects, including a high risk of
infection and organ injury. Because the side effects can be more
dangerous to your health than your diabetes, you're usually not
considered a candidate for transplantation unless your diabetes can't
be controlled or you're experiencing serious complications. On the
other hand, pancreas transplantation may be an option if you are age
45 or younger, have type 1 diabetes and need or have had a kidney
transplant, or if insulin doesn't control your blood sugar.
• Islet cell transplantation. Your pancreas contains about 1 million
islet cells, 75 percent to 80 percent of which produce insulin. The beta
cells that produce insulin reside in the islets. Although still considered
an experimental procedure, transplanting these cells may offer a less
invasive, less expensive and less risky option than a pancreas
transplant for people with diabetes. In islet cell transplantation,
doctors infuse fresh pancreas cells into the liver of the person with
diabetes. The cells spread throughout the liver and soon begin to
produce insulin. The liver, not the pancreas, is the site of the
transplant because it's easier to access the large portal vein in your
liver than it is to access a vein in your pancreas. What's more, cells
 that grow in the liver secrete insulin much like cells in the pancreas
do.

NATURAL METHODS OF TREAMENT FOR DIABETES

Hydrotherapy
It this natural method of treating diabetes, the water is used. It is one
dysfunctions. The solvent property of water and its ability to absorb and
studies have shown that regular uses of hydrotherapy provide lots of benefits
provides better and improved sleep, reduces blood sugar level in the body
also provides support to individuals who find exercise difficult. It is
diabetes. It is also known as hot tub therapy. Hence, Hydrotherapy is shapes.
Usually extremities are treated in these tanks.

Detoxification
It consists of the use of the short periods of fasting or controlled diets
and supports itself of toxic substances.

Mud therapy
Generally earth provides us with food, our main source of energy.
During element earth in the form mud or clay packs. Even mud baths are
uses food poisons from the body, for cooling the nerved system and for
activating vast the metabolic process of the entire body. When the digestive
system, and earth of impurities and toxemia in the body. Hence improving
elimination and re Mud bath treatment is used. There are many ways taking
mud treatment ordinary mud. The clay should be of such a consistency that
it could be body by loose cloths wrapping. The clay, in winter should be
sufficiently war.

Chromotherapy
Chromo therapy is a natural method of treatment of various
diseases, which their natural balance. Hence, we can say, it is a therapeutic
method, which spectrum with specific curative properties. By using the
property to region the harmony and order of the organism. The colour
illumination helps in physical well-being.

Medicinal plants

Herbal remedies from medicinal plants (Known in the Caribbean as

bush medicines) have been used traditionally in regions where access to
formal health care is limited. There are several reasons why use of bush
medicines should be studied: herbal remedies may have recognizable
therapeutic effects (Bailey CJ, Day C. et.al. 1989). They may also have
toxic side-effects (Keen RW. Deacon AC.et.al.1994) and use of bush
medicines provides an indication of beliefs about illness and its treatment
that may conflict with beliefs of workers in the formal health care system
(Morgan M, Watkins et.al.1998).
Over the last three decades chronic disorders such as diabetes and
hypertension have emerged as the major causes of adult morbidity and
mortality in the Caribbean islands. (Gulliford MC 1994). Successful control
of health problems stemming from diabetes and hypertension requires active
participation of patients in their own care, but as Morgan has pointed out,
‘health care providers’ limited understanding of Caribbean patients’
concepts of illness and ideas about treatment are obstacles to establishing
effective patterns of self care.
Plans are used to treat many ailments. India has about 45,000 plants
species and several thousands have been claimed to possess medicinal
properties. Medicinal plants used to treat hypoglycemic or hyperglycemic
conditions are of considerable interest for ethno-botanical community as
they are recognized to contain valuable medicinal properties in different
parts of the plant and a number of plants have shown varying degree of
hypoglycemic an anti-hyperglycemic activity. (Grover JK, et al. 2002). The
active principles if many plant species are isolated for direct use as drugs,
lead compounds or pharmacological agents (Fabricant DS, Famsworth NR,
et al. 2001). Several species of medicinal plants are used in the treatment of
diabetes mellitus, a disease affecting large number of people world-wide.
Traditional plant medicines or herbal formulations might offer a natural key
to unlock diabetic complications. (Nammi.S, et al.2003).

Diabetes mellitus is the major endocrine disorder (Burke JP, et al.

2003). Responsible for renal failure, blindness or diabetic cataract
(Thylefors. B1990). Poor control increased risk of cardiovascular disease
including atherosclerosis and AGE (advanced glycogen end) products.
Antioxidants play an important role to protect against damage by reactive
oxygen species and their role in has been evaluated. Many plant extracts and
products were shown to possess significant activity. (Sabu MC, Kuttan R, et
al. 2002).
 Since ancient times, plants have been an exemplary source of
medicine. Ayurveda and other Indian literature mention the use of plants in
treatment of various human ailments. India have about 45000plant species
and among them, several thousands have been claimed to possess medicinal
properties. Researches conducted in last few decades on plants mentioned in
ancient literature or used traditionally for diabetes have shown anti-diabetic
property. The present paper reviews 45 such plants and their products
(active, natural principles and crude extracts) that have been mentioned/used
in the Indian traditional system of medicine and have shown experimental or
clinical anti-diabetic activity. Indian plants which are most effective and the
most commonly studied in relation to diabetes and their complications are:
Allium cepa, Allium sativum aloe vera, Cajanus cajan, Coccinia indica,
Caesalpinia bonducella, Ficus bengalenesis, Gymnema sylvestre,
Momordica charantia, Ocimum sanctum, Pterocarpus marsupoim Swertia
chirayita, Syzigium cumini, Tinospora cordifolia and Tinospora cordifolia
and Trigonella foenum graecum. Among these we have evaluated
M.charantia, Eugenia jambolana, Mucuna puriens, T.cordifolia,
T.cordifolia,T.foenum graecum, O.sanctum, P.marsupuum, Murraya
koeingii and Brassica juncea. All plants have shown varying degree of
hypoglycemic and anti-hyperglycemic activity.

Cinnamomum zeylanicum
Cinnamomum (verum, synonym c. zeylanicum) is a small
evergreen tree belonging to the family Lauraeae, native to sri Lanka.
(Enzyclopaedia Britanica 2008). Or the spices obtained from tree’s
bark .
Scientific classification
Kingdom : Plantae
Division : Magnoliphita
Class : Magnoliopsida
Order : Laurales
Family : Lauraceae
Genus : cinnamomum
Species : C.verum

Binomial name
Cinnamomum verum
Synonyms
C. zeylanicum Blume

Nomenclature and taxonomy

The name cinnamon comes from Phoenician through the Green
Kinnamomum. The botanical name for the spice – Cinnamomum
Zeylanicum- is derived from Sri Lanka’s former name.

In Tamil - lavangapattai
In Malayalam - karugapatta
In Telugu - Dhalchi8na chalka (bark)
In Hindi - Dalchini
In English - Cinnamon

Applications of cinnamon
 Cinnamon bark is widely used as a spice. It is principally employed in
cookery as a condiment and flavoring material. It is used in the
preparation of chocolate, especially in Mexico, which is the main
importer of true cinnamon. (www.fao.org.) It is also used in the
preparation of some kinds of desserts, such as apple pie, donuts and
cinnamon buns as well as spicy candies, tea, hot cocoa and liqueurs.
True cinnamon, rather than cassia, is more suitable for use in sweet
dishes. In the Middle East, it is often used in savory dishes of chicken
and lamb. In the United States, cinnamon and sugar are often used to
flavor cereals, bread-based dishes and fruits, especially apples; a
cinnamon-sugar mixture is even sold separately for such purposes.
Cinnamon can also be used in pickling.
 Cinnamon bark is separately for such purposes. Cinnamon can also
be used in pickling. Cinnamon bark is one of the few spices that can
be consumed directly. Cinnamon powder has long been an important
spice in Persian cuisine, used in a variety of thick soups, drinks and
sweets. It is often mixed with rosewater or other spices to make a
cinnamon-based curry powder for stews or just sprinkled on sweets
treats.

 Its flavor is due to an aromatic essential oil that makes up 0.5% to 1%

of its composition. This oil is prepared by roughly pounding the bark,
macerating it in seawater, and then quickly distilling the whole. It is
of a golden-yellow color, with the characteristic odor of cinnamon and
a very hot aromatic taste. The pungent taste and scent come from
cinnamic cinnamon and a very hot aromatic taste. The pungent taste
 and scent come from cinnamic aldehyde or cinnamaldehyde (about
60% of the bark oil) and by the absorption of oxygen as it ages, it
darkens in color and develops resinous compounds. Other chemical
components of the essential oil include ethyl cinnamate, eygenol
(found mostly in the leaves), beta-caryophyllene, and linalool and
methyl chavico).

Cinnamomum zeylanicum medicinal Properties

 In medicine it acts like other volatile oils and once had a reputation as

cure for colds. It has also been used to treat diarrhea and other
problems of the digestive system. (Shan B, Cai YZ, et al 2005)
Cinnamon is high in antioxidant activity (Felter, Harvey, 2007) the
essential oil of cinnamon also has antimicrobial properties (Lopez P,
2005) which can aid in the preservation of certain foods.

 Cinnamon has been reported to have remarkable pharmacological

effects in the treatment of Type 2 diabetes mellitus and insulin

resistance. However, the plant material used in the study was mostly
from cassia and only few of them are truly from Cinnamomum
zeylanicum (see cassia’s medicinal uses for more information about
its health benefits). (Khan A, Safdar M. et al 2003). Recent
advancement in phytochemistry has shown that it is a cinnamtannin
B1 isolated from C.zeylanicum which is of therapeutic effect on Type
2 diabetes, (Taher, Muhammad et al.) with the exception of the
postmenopausal patients studied on C. cassia. (Kristol.
2008))Cinnamon has traditionally been used to treat toothache and
 fight bad breath and its regular use is believed to stave off common
cold and digestion.

 Cinnamon has been proposed for use as an insect repellent, although it

remains untested. Cinnamon leaf oil has been found to be very

efective3 in killing mosquito larvae.(Ranjbar, Akram et al 2008) The
compounds cinnamaldehyde, cinnamyl acetate, eugenol and anethole
that are contained in cinnamon leaf oil, were found to have the highest
effectiveness against mosquito larvae.

 It is reported that regularly drinking of Cinnamomum zeylanicum tea

made from the bark could be beneficial to oxidative stress related
illness in humans, as the plant part contains significant antioxidant
potential.

 Cinnamon may also be an aphrodisiac. (Nickell, Nancy L. et al 1998).

One teaspoon of Cinnamon contains as many antioxidants as a full

cup of pomegranate juice and ½ a cup of blueberries.

Scope of Present Investigation

Diabetes mellitus is characterized by an initial loss of glucose

homeostasis resulting from defects in insulin secretion and or insulin
action or leading to impaired metabolism of glucose and other energy
–yielding metabolites. It is also accompanied by hyperglycemia,
dyslipideamis, hypertention, decreased fibronolytic activity, increased
platelet aggregation and severe atherosclerosis, all of which are
potential risk factors.
 Through, several drugs targeting carbohydrate hydrolyzing
enzymes (Pseudosacchrides), release of insulin from pancreatic β-cells
(sulphonylurea), glucose utilization (biuguanides), insulin
sensitization and PPAR γ agonists (glitazones) are in clinical practice,
there is a growing market for antidiabetic agents. Many of these oral
antidiabetic agents have been reported to show serious adverse effects.
The multifactorial etiology and multiple pathogenic manifestation of
diabetes demands multi-model therapeutic approach and future
therapeutic strategies require evaluation of efficiency of combination
of drugs. Management of Diabetes sans side effects is still a
challenge for the pharmaceutical world.

Traditional knowledge of medicinal plants has become a

recognized tool in research for the search of new sources of drugs and
neutraceuticals. In fact, metformin, one of the most prescribed
glucose –lowering medicines currently used, is derived from a
chemical isolated from a plant. The World Health Organization has
infact recommended the use of herbal medicine especially in
developing countries (WHO, 2002). Because of their perceived
effectiveness, minimal side effects in clinical experience and
effectiveness, herbal drugs are prescribed widely even when
biologically active compounds are not known.

Cinnamomum zeylanicum extracts are used in folk treatments

of diabetes. This natural remedy has not received scientific of
medical scruting and was not listed in the most recent and extensive
reviews on traditional plant medicines for treatment of diabetes. The
present study was undertaken to investigate the effect of the oral
 administration of an aqueous extract of Cinnamomum zeylanicum on
normal and alloxan diabetic rats, as an approximation to the possible
mechanism of action

Animals

Rats used in this experiment were highly inbred male Wister

Albino rats from laboratory (APCAS). The rats weighed 160g were used.
The animals were housed in specious caged under hygienic conditions (13h
light 12h dark cycle at room temperature) and maintained on commercial
pellet diet containing protein- 21%, lipid -5%, nitrogen free extract -55%
and provided with metabolically energy at 3600Kcal/kg and also enriched by
vitamins and minerals. It was supplied by the “Hindustan Lever Limited”
Mumbai marked under the trade name “Gold mohur feeds” water was
provided. The rats were kept in animal’s house for ten days before starting
the experiments.

Induction of diabetes mellitus

Alloxan monohydrate induced diabetes mellitus was produced in a

batch of hypogenic male albino rats by injecting intrapreophrea; cavity a
single dose (40mg/kg weight) of 2% alloxan monohydrates solution in
saline, after thus have been fasted for 24hours. This single dose of alloxan
produced persistence hyperglycemia after 7 days it was observed that the
condition for 5 days. The animals showed the following signs of the
condition: polydipsia (abnormal thirst), polyuria (increased urine volume),
weight loss (due to lean mass loss), asthenia weakness (due to the inability
to use glucose as a source of energy), dehydration (due to the animal body’s
attempts to get rid of the excess blood glucose as the normal process of
storing glucose in the body cells is impair

In order to asses the effect of alloxan and to chemically

establish the diabetic condition, an incision was done in any of the
four veins in the tail of the rat 7 days after induction. After 7 days
start (or) behind the treatment to the rats (Oi K, Komori, H, 1997,
Machado, JLM, Godoy, p, 1981, Dunn, J.S., 1943, Calva, K.C.,
2001).

Preparation of plant material: Bark

Cinnamomum zeylanicum (bark) was purchased from the
local market at Thiruvannamalai. The bark was dried and finely
powdered in a mechanical mixer. 10g of finely powdered cinnamon
was weighed mixed with 100 ml of water and kept in a water bath at
60◦C for two hours and filtered . This extract was diluted with water
(1:10) and was administered orally to rats.( Singapore Med J 2006)

Experimental Design
Experimental animals are divided into five treatment groups.

Group- I
Normal
Six albino rats are maintained in normal condition.
Group – II
Diabetic
The rats were made diabetic by administration of 2% Alloxan
monohydrate. The rats were fasted for 16 hours but had been allowed free
access to water. Alloxan monohydrate was dissolved in sterile normal saline
immediately before use and injected intraperinotical cavity in a dose 2%
Alloxan monohydrate solution in saline. The single dose of alloxan
produces persistence hyperglycemia after 24 hours and it was observed that
the condition was maintained for 6 days.

Group –III
Cinnamomum zeylanicum control

The rats were in normal condition control rats also maintain same
procedure control rats receiving cinnamomum zeylanicum extract. 160
mg/kg body weight of rats. This herbal dissolved in 20ml of water and orally
administered for every 24 hours for 20 days

Group –IV
Treatment diabetic
Alloxan diabetic rats receiving cinnamomum zeylanicum extract
2ml per day orally administrated for every 24 hours for 20 days.

Group – V
Glibenclamide Treatment
 Diabetic rats given with glibenclamide (600microgram/kg body
weight) in aqueous solution daily introgastric tube for 20 days.
The animal were dosed through provided the incubation everyday
before any food was given. Food and water were provided the duration of
treatment was 20 days. After the treatment period, the rats were sacrificed,
and blood was drawn from ventricles and serum separated for various
biochemical estimations. On the final day of experiment, liver tissue was
collected. Liver was excised from each animal; the tissue was washed with
ice cold saline and homogenized in Tris HCL buffer pH-7.5. The serum
obtained was used immediately for the estimation of blood glucose, total
serum protein, serum cholesterol, serum triglycerides, glycosylated
hemoglobin, urea, and insulin. Liver tissue was used for assay the enzyme
activated of Glutamate Pyruvate Transaminase (GPT), Hexokinase, glucose-
6 phosphatase and fructose 1, 6 bis phosphatase. The glycogen obtained after
over nights precipitation was then estimated.

ESTIMATION OF BLOOD GLUCOSE

Blood glucose was estimated by the method of Sasaki and

Matsui (1972).

Reagents

1. Trichloro acetic acid : 10%

2. O-toluidine reagent : 12.5 g of thiourea and 12.0 g boric acid were

dissolved in 50ml of water by heating over a mild flame. 75 ml of

redistilled O-toluidine and 375 ml of acetic acid were mixed with

 thiourea – boric acid mixture and the total volume was upto 500ml

with water. The reagent was left in refrigeration overnight and

filtered.

3. Standard glucose solution : 10mg of pure glucose was dissolved in

100ml of 0.2% boric acid in water.

4.Procedure

0.1ml of blood was mixed with 1.9ml of TCA solution to

precipitate protein and then centrifuged. 1.0ml of the supernatant was mixed

with 4 ml of O-toluidine reagent and was kept in a boiling water bath for

about 15 min. and the green colour developed was read at 600 nm in a

Shimadzu spectrophotometer. A set of standard glucose solutions were also

treated similarly.

The values were expressed as mg/dl blood.

ESTIMATION OF PLASMA PROTEIN

The protein was estimated according to the method of

Lowry(1951).

Reagents

1. Alkaline copper reagent

Solution A : 2% sodium carbonate in 0.1N sodium hydroxide solution.

 Solution B : 0.5% copper sulphate in 1% sodium potassium tartarate.

50ml of solution A and 1 ml of solution B were mixed just before use.

2. Folin’s – Phenol reagent

100 g of sodium tungstate, 25g of sodium molybdate, and 700 ml of

distilled water, 50ml of 85% O-phosphoric acid and of concentrated

hydrochloric acid were added. The mixture was refluxed gently for

about 10h. After cooling, 150g of lithium sulphate, 50ml of water and

a few drops of bromine were added. The mixture was boiled for

15min. without a condenser to remove excess bromine. The contents

were cooled, made upto 1 litre and then filtered. The above stock

solution was diluted 1 : 2 with water prior to use.

3. Standard protein solution

10mg of crystalline Bovine serum albumin was dissolved in

100ml of distilled water.

Procedure

A serum was mixed with 4.5ml of alkaline copper reagent and then

allowed to stand at room temperature for 10min. Then 0.5ml of Folin’s

phenol reagent was then added. The blue colour developed was read at

640nm in a shimadzu spectrophotometer after 10min. A standard graph was

obtained using bovine serum albumin solution.

 The protein values were expressed as g/dl.

ASSAY OF INSULIN BY ELISA

Principle

The Insulin ELISA is a solid phase Enzyme linked immuno sorbent

assay performed on micro titre plate. It is based on the oligoclonal system in

which several monoclonal antibodies (Mabs) directed epitopes of insulin are

used. The use of several distinct Mabs avoids hyperspecificity and allows

high sensitive assays with extended standard range and short incubation

time.

Standards or samples containing insulin (INS) react with capture

antibodies (Mabs 1) coated on a plastic well and with monoclonal antibodies

(Mabs 2) labelled with horse raddish perxoidase (HRP). After an incubation

period allowing the formation of a sandwich coated Mabs 1 –INS- Mabs 2 –

HRP, the microtitre plate is washed to remove unbound enzyme labelled

antibodies.

The substrate solution tetramethylbenzydine (TMB – H 2O2) is added

and incubated. The reaction is stopped with H2SO4 and the microtitre plate is

read at the appropriate wavelength. The amount of substrate turnover is

determined colorimetrically by measuring the absorbance which is

proportional to the insulin concentration.

MATERIALS REQUIRED:

1. Microwell strips (96 wells) :

Monoclonal anti insulin Ab coated wells 8 x 12 strips.

2. Enzyme Conjugate (11 ml) Anti-insulin Ab conjugate to horse

raddish peroxidase.

3. Reference Standard Set (0.75 ML /each)

Human insulin references:

0, 5, 25, 50,100,200 IU/ml calibrated against first who IRP of

insulin.

4. Solution-A

Buffer solution containing H2O2 .

5. Solution-B

Tetra methyl benzidine (TMB)

6. Concentrated Wash Buffer (20x) – 50ml

7. Well holder for securing individual well

Specimen collection and preparation

 1. Serum or plasma may be used for the test, while preparing serum

samples remove the serum from the clot as possible to avoid

haemolysis.

2. Specimen should be free from microbial contamination and may be

stored at 2-8°C f or a week.

Sample preparation

Tube Dilution

The serum sample was diluted with the diluted buffer (in 11

dilution).Separate tips were used for each sample and then discarded as bio

hazardous waste.

Microwell dilution

About 100 ml of same buffer was added to microwell followed by

10µ l of serum sample to be tested.

Preparation of wash buffer

The 25X wash buffer was diluted to 1X concentration and the

required volume of wash buffer (50 ml per each strip) was prepared.

Preparation of working conjugate

 The 100X conjugate was diluted into 1X concentration at the

required volume approximately 1ml per strip.

Preparation of working subtrate conjugate

The 100X TMB

Concentrate was diluted to 1X concentration at the required volume

of 1ml per strip.

TEST PROCEDURE

1. Required number of Micro ELISA STRIP was fixed to the Micro

ELISA strip holder sequentially.

2. 100µ l of the sample dilution buffer was added into A-I well as a

blank, 100 µ l of the negative control was added into B-I and C-I

respectively. 100 µ l of the positive control was added into D-I, E-

I, F-I respectively.

3. In the following wells 100 µ l of the sample dilution buffer was

added followed by 100 µ l of serum.

4. The pellets were sealed with cover and incubated at 37°C ± 2°C for

30 minutes.
 5. The plates were taken out from the incubator after the incubation

time was over and washed five times with 1X washing solution.

6. 100 µ l of the working conjugate solution was added to all the

wells

7. The plate was sealed and incubated at 34°C± 2°C for 30 minutes.

8. After incubation the conjugate was aspirated and washed 5 times

with 1X wash buffer.

9. 100 µ l of working substrate solution was added to each well and

incubated at room temperature for 30 minutes in dark.

10. After incubation 50 µ l of the stop solution was added. Then plates

were read at 450 nm using ELISA reader with 30 minutes.

The values were expressed as µ /ml blood.

ESTIMATION OF BLOOD UREA

Urea was determined in the blood by the method of Natelson (1951).

Reagents

1. Diacetylmonoxine reagent : 2g of diacetylmonoxine was dissolved in

100ml of 2% acetic acid.

 2. Sulphuric acid – phosphoric acid mixture : 25ml of concentrated

sulphuric acid, 75ml of 85% O-phosphoric acid and 70ml of distilled

water were mixed.

3. 10% sodium tungstate solution

4. 0.67N sulphuric acid

5. Standard urea : 20mg of urea was dissolved in 100ml of distilled

water

Procedure

0.1ml of blood was added to 3.3ml of water and mixed with 0.3ml of

10% sodium tungstate and 0.3ml of 0.67N sulphuric acid reagents. The

suspension were centrifuged and to 1.0ml of the supernatant, 1.0ml of water,

0.4ml of diacetyl monoxime and 2.6ml of sulphuric acid – phosphoric acid

reagents were added in that order and kept in a boiling water bath for 30

min. cooled and the colour developed was measured at 480nm in a shimadzu

spectrophotometer. Aliquots of standard urea were also treated in a similar

manner.

The values were expressed as mg/dl blood.

ESTIMATION OF CHOLESTEROL

Cholesterol was determined in the blood by the method of Parekh and

Jung method (1970) using the Ferric sulphate colour reaction in the presence

of uranyl acetate.

Reagents

1. Ferric-Uranyl acetate reagent

0.1ml of water and 3.0ml of concentrated ammonia were added to

500mg of crystalline ferric chloride. The precipitate was washed

several times with distilled water to get rid of ammonia and was

dissolved in cholesterol grade glacial acetic acid and made upto one

litre with acetic acid. 100mg of uranyl acetate was then added to it

and the contents were well shaken and left overnight. The reagent

was stored in a brown bottle.

2. Sulphuric acid – Ferrous sulphate reagent

To 100ml of cholesterol grade glacial acetic acid 100mg of anhydrous

ferrous sulphate was added and shaken well. To this 100ml of

concentrated sulphuric acid was added. After cooling, the volume

was made upto 1 litre with concentrated sulphuric acid.

3. Cholesterol standard
 Reagent grade cholesterol was recrystallised from ethanol. The stock

standard was prepared by dissolving 200mg of cholesterol in 100ml

of cholesterol glacial acetic acid.

Procedure

10ml of ferric-uranyl acetate reagent was added to 0.1ml of plasma

sample, mixed well allowed to stand for 5 minutes and centrifuged. 3.0ml of

the supernatant was taken for analysis. Similarly, 0.1ml of standard

cholesterol was mixed, and 3.0ml aliquots were taken. Blank tube contained

3.0ml of ferric acetate-uranyl acetate reagent. 2.0ml of sulphuric acid –

ferrous sulphate reagent was added to all the tubes and mixed well. The

colour intensity was measured at 560nm after 20 minutes in a shimadzu

spectrophotometry.

The values were expressed as mg/dl plasma.

ESTIMATION OF TRIGLYCERIDES

Triglycerides were estimated by the method of Foster and Dunn

(1973). Triglycerides are extracted by isopropanol, which upon

saponification with potassium hydroxide yield glycol and soap. The

glycerol liberated is treated with metaperiodate, which release

formaldehyde, formic acid and iodide. The formaldehyde released reacts

with acetylacetone and ammonia forming yellow coloured compound, the

intensity of which is measured at 405 nm.

Reagents

1. Isopropanol

2. Activated aluminium oxide (Neutral)

3. Saponification reagent: 5.0 g of potassium hydroxide was dissolved in

60ml of distilled water and 40ml of isopropanol was added to it.

4. Sodium metaperiodate reagent: 77g of anhydrous ammonium acetate

was dissolved in about 700ml of distilled water. 60ml glacial acetic

acid was added to it followed by 650mg of sodium meta periodate.

The mixture was diluted to 1.0 litre with distilled water.

5. Acetyl Acetone reagent: 0.75ml of acetyl acetone was dissolved in

60ml of distilled water and 40ml of isopropanol was added to it.

6. Standard triolein solution: 1g of triolein was dissolved in 100ml of

isopropanol. 1.0ml of stick standard was diluted to 100ml to prepare a

working standard of concentrated 100µ g of triolein/ml.

Procedure
 To an aliquot of serum / lipid extract, evaporated dryness, 0.1ml of

methanol was added followed by 4.0ml of isopropanol. 0.4g of Alumina

was added to all tubes and shaken well for 15 minutes. Centrifuged another

2.0ml of the supernatant fluid was transferred to labelled tubes. The tubes

were placed in a water bath at 650 for 15 minutes for saponification after

adding 0.6ml of the saponification reagent followed by 1ml of metaperiodate

reagent and 0.5ml of acetyl acetone reagent, after mixing. The tubes were

kept in a waterbath at 650C for one hour, the contents were cooled and

absorbance was read at 405nm. A serious of standards of concentrated 8 to

40µ g triolein was treated similarly along with a blank containing only the

reagents. All the tubes were cooled and read at 405nm. The values were

expressed as mg/dl serum.

 TEST FOR GLYCOSYLATED HAEMOGLOBIN (HbA1C)

(Gabby, K.H. et al.) (Gonen, B, 1978)

Principle

A hemolysed preparation of whole blood is mixed continuously for

five minutes with a weakly binding cation exchange resin. The labile

fraction is eliminated during the hemolysate preparation and during the

binding. During this mixing non glycosylated hemoglobin binds to the ion

exchange resin leaving HbA1C free in the supernatant. After the mixing

period, a filter separator is used to remove the resin from the supernatant.

The percent glycosylated hemoglobin is determined by measuring

absorbance of GHb fraction and total haemoglobin (THb) fraction.

Reagents

1. Lysing Reagent

2. Ion exchange resin

3. Control Lyophilized

4. Resin separators

Precautions

Glycosylated haemoglobin kit is for in vitro diagnostic use only

  Do not mix or use the reagents from test units that have different lot

numbers

 Bring reagents to assay temperature before use. Ensure constant

assay temperature (preferably 230C) of resin during GHb separation

of the assay

 Initial use of the control is advised to check test system performance

within assay limits

Procedure

Step I: Hemolysate preparation

Pipette 0.25ml of lysing reagent in a test tube. Then add 0.005ml of

well mixed sample. Mixed well the sample and allowed to stand at room

temperature for 15 minutes.

Step II: GHb separation and assay

Bring a resin tube to assay temperature by incubating the tube in a

water bath. Added 0.1ml of hemolysate to it. Place a resin separator in the

tube, so that the rubber sleeve was at the upper mark of the tube (3 cm above

the resin level). Contents are mixed on vortex mixer continuously for 15

minutes. Allowed push down the resin separator in the tube until the resin
was firmly packed. Poured the supernatant directly into a cuvette and

measured the absorbance against distilled water.

Step III: Total Hemoglobin (THb) Assay

Pipette 5ml of deionized water into a test tube. Then add 0.02ml of

hemolysate (from step I). Mixed well and read absorbance against distilled

water.

Normal values:

Whole blood - 6.0 – 8.7%

Calculations:

Absorbance of GHb

GHb % = x 10 x Temperature factor (Tf)

Absorbance of THb

For assay at 230C Tf = 1.0; at 300C Tf = 0.9

ESTIMATION OF LIVER GLYCOGEN

The extraction was carried out by the method of Morales (1973).

Glycogen was precipitated from the alkali extract of the tissues by

adding 1 : 3 volume of 95% ethanol and a drop of 1 M ammonium acetate.

The tubes were kept in boiling water bath for 5 minutes. After cooling, the
tubes were shaken well and placed in a freezer overnight. The precipitated

glycogen was then collected by centrifugation at 3000g for 40 minutes. The

precipitated was dissolved in water, reprecipitated with Alcohol and

centrifuged again. The final precipitate was dissolved in 3.0ml of water and

heated for 5 minutes in boiling water bath and different aliquots were used

for the estimation of glycogen.

Procedure

Suitable aliquots of glycogen were made upto 1.0ml with water. A set

of standard glucose solutions (25 - 100µ g) and blank containing water

alone were setup. All the tubes were cooled in an ice bath and 4.0ml of

anthrone reagent was added. The contents of the tube were mixed well. All

the tubes were covered with glass marbles and heated for 20 minutes in

boiling water bath. The tubes were cooled and the green colour developed

was read at 640nm in a Shimadzu spectrophotometry.

The values were expressed as mg of glucose / g of wet tissue.

ASSAY OF GLUTAMATE PYRUVATE TRANSAMINASE (GPT)

The enzyme activity was assayed by the method of Mohur and Cook

(1957).

Reagents

1. Buffered substrate : (0.1M Phosphate buffer, pH 7.4)

1.5g of Dipotassium hydrogen phosphate, 0.2g of Potassium

dihydrogen phosphate, 0.03g of 2-Oxoglutaric acid and 1.78g of DL-

alanine were dissolved in distilled water. The pH was adjusted to 7.4

and made upto 100ml

2. 200mg of 2, 4-dinitrophenyl hydrazine (DNPH) in 100ml of 2N

Hydrochloric acid.

3. 0.4N Sodium hydroxide

4. Standard Pyruvate

Procedure

To 1.0ml of buffered substrate, 0.1ml of serum or 0.2ml of tissue

homogenate was added and incubated at 370C for exactly 30 min. The

reaction was arrested by adding 1.0ml of DNPH and left aside for 20min at

room temperature. Colour developed by the addition of 10ml of 0.4N

sodium hydroxide was read at 540nm.

 The enzyme activity in serum was expressed as IU/L. The enzyme

activity of the tissue was expressed as n mol of pyruvate formed / h / mg

protein.

ASSAY OF HEXOKINASE

Hexokinase (ATP : D-hexose-6-phosphotransferase) was assayed by

the method of Brandstrup (1957).

Reagents

1. 0.005 M Glucose solution

2. 0.72 M ATP solution

3. 0.05 M Magnesium chloride solution

4. 0.0125 M Dipotassium hydrogen phosphate solution

5. 0.1M Potassium chloride solution

6. 0.5 Sodium fluoride solution

7. 0.01 M Tris-HCl buffer (pH 8.0).

Procedure

The reaction mixture in a total volume of 5.0ml contained the

following: 1.0 ml of glucose solution, 0.5ml of ATP solution, 0.1ml of

magnesium chloride, 0.4ml of dipotassium hydrogen phosphate solution,

0.4ml of potassium chloride, 0.1 ml of sodium fluoride solution and 2.5ml of

Tris – HCl buffer (pH 8.0). The mixture was preincubated at 370C for 5

min. The reaction was initiated by the addition of 2.0ml tissue homogenate.

1.0ml aliquot of the reaction mixture was taken immediately (zero time) to

tubes containing 1.0ml of 10% TCA. A second aliquot was removed after

30 minutes of incubation at 370C. The precipitated protein was estimated by

the O-toluidine method of Sasaki and Matsui (106) as described previously.

A reagent blank was run with each test.

The difference between the two values gave the amount of glucose

phosphorylated. The enzyme activity was expressed as n moles of glucose-

6-phosphate formed/h/mg protein.

ASSAY OF GLUCOSE-6-PHOSPHATASE

Glucose-6-phosphatase was assayed according to the method of King

(1965).

Reagents

1. 0.1M Citrate buffer (pH 6.5)

2. Substrate : Glucose-6-phosphate, 0.01M in distilled water

3. 10% TCA
Procedure

The incubation mixture in a total volume of 1.0ml contained 0.3ml of

buffer, 0.5ml of substrate and 0.2ml of enzyme extract. Incubation was

carried out at 37°C for 60 minutes. The reaction was terminated by the

addition of 1.0ml of 10% TCA. The suspension was centrifuged and the

phosphorus content of the supernatant was estimated according to the

method described by Fiske and Subbarow.

The enzyme activity was expressed as moles of phosphorus

liberated/mg protein under the incubation conditions.

ASSAY OF FRUCTOSE-1, 6-BIPHOSPHATASE

Fructose-1, 6-biphosphatase was assayed by the method of Gancedo

and Gancedo (1971).

Reagents

1. Tris-HCl buffer : 0.1M (pH 7.0)

2. Substrate : Fructose-1, 6-biphosphatase – 0.05M solution

3. Magnesium chloride – 0.1 M solution

4. Potassium chloride – 0.1 M solution

5. EDTA solution – 0.001 M solution

6. Trichloroacetic acid : 10% solution

 7. Molybdic acid

2.5% ammonium molybdate in 3N sulphuric acid

8. Amino naphthol sulphonic acid (ANSA)

500mg of Amino naphthol sulphonic acid was dissolved in

195ml of 15% sodium bisulphate solution and 5ml of 20% sodium

sulphite solution was added for complete solubilization. The solution

was filtered and stored at 40C in a brown bottle.

9. Phosphorous stock standard

35.1 mg of potassium dihydrogen phosphate was dissolved in 100ml

of distilled water (80 µ g/ml).

Procedure

The assay medium in a final volume of 20ml contained 1.0ml buffer,

0.4ml of substrate, 0.2ml each of magnesium chloride and potassium

chloride, 0.1ml of EDTA and 0.1ml of the enzyme source. The incubation

was carried out at 37°C for 15 minutes. The reaction was terminated by the

addition of 1.0ml of 10% TCA. The suspension was centrifuged and the

phosphorus content of supernatant was estimated according to the method

described by Fiske and Subbarow. To the aliquot of the supernatant, 4.1ml

of distilled water and 0.5ml of ammonium molybdate were added. After 10

minutes 0.2ml of ANSA was added. The tubes were shaken well, kept aside

for 20 minutes and the blue colour developed was read at 620nm against a

water blank in a Shimadzu spectrophotometer.

Enzyme activity was expressed as n moles of phosphate liberated/mg

protein.

Statistical Analysis

The different of biochemical parameters were measured using

the statistical method of Analysis of Variance (ANOVA). Analysis of

variance refers to the examination of differences among the samples. It is an

extremely useful technique concerning research in biology. It is an

abbreviated as ANOVA (Analysis of Variance AN + O + VA). It is a

statistical technique specially designed to test whether the means of more

than the quantities population are equal.

The statistical significance was assessed using one-way

Analysis of Variance (ANOVA) using SPSS 12.0 version (SPSS, Cary, NC,

USA) followed by Bonferroni’s multiple comparison test (BMCT). The

values are expressed as mean ± SD and P < 0.05 was considered significant.
Result and discussion

Diabetic is a group of metabolic disease characterized by

hyperglycemia high blood sugar level. Non-insulin dependent diabetes
mellitus is the commenset form of form of diabetes globerties as well as
in India. Hereditary factor obesity sedentary life style and aging have been
shown to increase the risk for Diabetes. The proper medical care and a
regular monitoring of Diabetes is essential not only to keep the diseases and
the control. But also to prevent an assortment of other Diabetes related
problems because no where cure has been identify hince, management of
Diabetes with diet exercise and drug has been enabasize.
Anti diabetic drugs treat diabetes mellitus by lowering blood
glucose levels in the blood with the exceptions of insulin. All the
administered orally , and are also called oral hypoglycemic agent , herbs for
diabetes are used more and more to compliment or some times replace
conventional diabetic drugs. It has been reported that cinnamomum has
insulin like activity and it contains like activity and it contains and active
ingredient water soluble polyphenolic compound. It initiate insulin triggers
its receptor and work synergistically with insulin Cinnamon also have anti
lipidimic effect (Jarvill and Karjee et al 2003).
The present study was conducted to find out the effect of orally
administration of cinnamomum zeylanicum on normal and alloxan diabetic
as approximation to the possible mechanism of action diabetes mellitus
was induced in albino rats by injecting alloxan monohydrate inraperitonial
cavity a single dose of 40mg/ kg of body of 2% alloxan anhydrate solution
in saline . After these have been fasting for 24 hours hyperglycemia has
been produced after a one week it was observed that a condition was
minutes for 5 days . Animal were dosed through every day before food and
water provide 20 days. After the treatment period the rats were sacrifice
blood was drawn from ventricle and serum separated for various
biochemical estimation.

The serum protein, serum cholesterol, serum triglycerides,

glycosylated haemoglobin, urea and liver tissue was use for assay of enzyme
activities of glutamate pyruvate transaminase (GPT), Hexokinase, glucose-6-
phosphatase and fructose-1, 6 biphosphatase. The glycogen obtained after
overnight prescribed was then estimated to bring out positive conclusion the
result are discuss with available data’s describe below.
After the induction of diabetes by injecting freshly prepares alloxan
through intraperitoneal cavity, it was confirmed by testing of glucosuria in
the urine using glucose indicator sticks, Diabetes induced within 7 days.
The changes in the body weight of different experimental groups were
shown in fig 1.

The body weight of the alloxan induced diabetic group II rats was
found to be reduced. On treatment with Cinnamomum zeylanicum and
Glibenclamide on Group IV and V, the body weight was gained comparing
to the normal and control rats of Group I and III. This shows Cinnamomum
zeylanicum exhibited considerable gain of body weight.

Fig 2 depicts the amount of food in grams and volume of fluid in ml

taken by the rats. The value was increased in group 2 diabetic rats
comparing on treatment with normal and control rat of group I and III
Cinnamon and Glibenclamide the amount of food and volume of was
reverted back to the normal and treatment group of IV and V. increased fluid
in take and food is the one of the symptoms of Diabetes which has been
normalize on treatment effect of herbal.

The table 1 and fig 3,4,5 shows the level of blood glucose, serum
protein, and plasma insulin of different group of rats. Animal treated with
alloxan induced diabetes group II shows a significant elevated in blood
glucose when compared to group I and III of normal, control treated with
Cinnamomum zeylanicum extract. Alloxan , a β- islet cell cytotoxin ,
destroying the pancreatic β-cell leads to reduced secretion of insulin by the
pancreatic islet cell (Colca et al., 1983). Cinnamomum zeylanicum treated
group IV diabetic rats might enhance glucose utilization because of
significantly reduces the blood glucose levels in treated rats, This might be
due to restoration of delays insulin response or inhibition of intestinal
absorption of glucose due to reduction in the activity of indestinal
glycosidases like sucrase, maltase and lactase in the small intestine in the
cinnamon treated rat.(Sung Hg; kim , et al 2005) The similar reaction
might carried out in group V treated with Glibenclamide . There was a
marked reduction in the plasma protein content of untreated diabetic rats
group II when compared to that of normal and control rats of group I and III.
On administration of Cinnamomum zeylanicum extract to diabetic rats
restore the protein level almost equal to group V treated with Glibenclamide.
It might be due to the increased uptake or glucose by the cell by stimulating
the insulin receptor(IRS) I. (Mohamed H .Mahfouz et al 2010 ) This may
inhibit the protein catabolism leads to positive nitrogen balance(Miura and
kako.1995)
 The diabetic rat shows a significant decrease in plasma insulin. On
treatment with an extract to Cinnamon diabetic rat group IV restore the
plasma insulin significantly compares to Glibeclamide treated group V. the
treatment of cinnamomum zeylanicum extract to normal control rat of group
III did not show significant effect of plasma insulin from the existing beta
cells of pancreas. Anti diabetic effect of cinnamon might be due to the
increase insulin like activity of cinnamon.(Richa Soni and vibha Bhatnagar
2009)

Table 2 and Fig 6,7,8 depicts the level of serum, cholesterol and
triglycerides. Urea, cholesterol and triglycerides levels which has increased
after induction of diabetes was found to be decreased in group IV after
treatment with Cinnamomum zeylanicum extract almost equal to
Glibenclamide treated group V rats. There are no any significant changes in
the level of urea, cholesterol and triglycerides in the administration of
Cinnamomum zeylanicum of group III rats. High cholesterol level
associated with coronary heart disease (CHD) observed in diabetic patients.
The occurrence of coronary heart disease in assay of liver enzyme GPT
indicated a decrease in their activity. It may due to the Cinnamoum
zeylanicum extract in the treatment rats stimulate the pancreatic insulin
mimetic effect. which is the inhibitor of lipolytic action .The
Phytochemical analysis shows that flavanoid present in the cinnamon
methyl hydroxyl chalcone which activate the insulin action and reduces the
lipolytic action(khar et al) . Urea is the catabolic product of protein.
elevated urea nitrogen in diabetes may be due to enhanced catabolism of
both liver and plasma proteins (Almal, vistrup et al). After the treatment
with Cinnamomum zeylanicum in group IV treated rat, it was restores to
normal compares with glibenclamide treated group V. It shows the herbal
may stimulate the glucose metabolism.
The level of glycogen in blood, GPT in liver tissue and glycosylated
haemoglobin were represented in table 3 and fig(9,10,11). Alloxan
administration resulted in significant fall of glycogen level in group II
diabetic animals when compares to normal. On treatment with
Cinnamomum zeylanicum and glibenclamide shows an elevated in the
stored glycogen level.
Liver glycogen may be considered as the best marker for assessing
hypoglycemic activity of any drug. This indicates that the peripheral free
glucose is being stores in liver in the form of glycogen by improving
glycogenesis.(Saltiela.et al 1986) The fall of liver glycogen content
observed in diabetic rats may be due to lack of insulin in the diabetic state,
result in the inactivation of liver glycogen synthase activity.

The activity of liver GPT has been increased under hepatocellular

damage. It also increased spontaneously in diabetes (Poglioro and
Notarbartola.1961). On treatment with cinnamoum zeylanicum and
glineclamide in group IV and V, improvements were noticed in the level of
GPT indicates normalize insulin action and a restoration of normal
functioning of level .
They flavonoid present in the 1,3 diphynyl 2,propane 1-one . as the
insulin diphynyl mimitric effect by stimulating the receptor level and also
activate the glycogen syntase enzyme might be the case for rising are
glycogen in the group IV rats. (Jamal, WH Hawsaw et, al 2009)
HbA1C in diabetic induced group II rat has been restore to normal in
group treated rats comparable with Glibenclamide treated rats . There was
any significant in the levels of HbA1C control group III. The lower insuli
hyper glycemia in diabetic rat is well reflected in the absorbed in
glycosylated haemoglobin and decreased haemoglobin content is quite
suggestive of glycosylated of pH, a series issue of diabetes . That is
responsible for many secondary complications. (Ansarullah, et al 2009) and
treatment with cinnamomum its insulin mimetic effect may levels to
decreases in the level of glycosylated haemoglobin (Kannappan. S
Jayaraman et al. 2006)

The changes in the activity of hepatic hexokinase, glucose-6-

phosphatase and fructose 1, 6 bisphosphatase of different group were shown
in table 4 and depicted in fig. (12,13,14) Liver has the vital organ for glucose
metabolism. Glycolysis and gluconeogensis are the two prime
complementary events balancing the glucose level in our body. A marked
reduction in the activity of hexokinase was observed in diabetic animals.
The activity of liver glycogenic enzymes was elevated in diabetic rats. On
treatment they reversed the changes in the activity of hepatic enzymes. On
administration of cinnamomum zeylanicum to normal rats showed no
significant effect on hepatic enzymes when compares to other.

In experimental diabetic rats under persistent hyperglycemia, there is

a changes in the enzymes of glucose metabolism and lead to the
pathogenesis of diabetic complication especially neuropathy and micro
vascular disease® . One of the key enzymes in the catabolism of glucose is
hexokinase, which phosphorylates glucose to glucose-6-phosphatase.
Administration of herbal to alloxan induced rats resulted in an increased
activity of hexokinase which leads to increased utilization of glucose for
energy production through glycolysis ®. The activity of glucose 6-
phosphatase and fructose 1,6bisphosphatase are the regulatory enzymes in
the gluconeogenic pathway. Administration of Cinnamomum zeylanicum
significantly reduces the activities of Gluconeogenic enzymes in diabetic
rats. The level of plasma insulin was found to be increased significantly in
diabetic rats treated with herbal may be consequence for the reduction in the
level of gluconeogenic enzymes. The reduction in the activities of
gluconeogenic enzymes can result in the decreased concentration of glucose
in blood (Singapore Med J 2006)
Summary

Diabetes mellitus is a group of metabolic disease

characterized by hyperglycemia –high blood sugar levels which
results from defects in insulin secretion. The male albino rats were
induced diabetes by the intraperitoneal injection of Alloxan. After
the Cinnamomum zeylanicum treatment to diabetic rats the blood
glucose level returned back to near normal level. During diabetes
condition the amount of food and volume of fluid intake is high
when compared to the normal rats. The body weight is decreased
in diabetic rats when compared to normal rats. After the
Cinnamomum zeylanicum treatment these levels were back to near
normal level.

Animal treated with alloxan induced diabetes shows a

significant increase in blood glucose when compares to normal
rats. Changes were observed in the levels of serum protein and
insulin in diabetic condition. After the administration of
Cinnamomum zeylanicum , these level are corrected to near
normal level due to Cinnamoum zeylanicum might be increases
the release of insulin from the existing β-cells of pancreas. The
levels of urea, cholesterol and Triglycerides were increased after
induction of diabetes. After the administration of Ciannamomun
zeylanicum these levels were reverted back to near normal level.
Alloxan administration resulted in significant reduction of
glycogen level in diabetic rats. On treatment with Cinnamomum
zeylanicum, it shows an elevation in the stores glycogen level.

Changes were observed in the level of glucosylated

haemoglobin in diabetic rats. After the administration of
Cinnamomum zeylanicum, the level was restored to near normal
level. Altered activities of hepatic enzymes such as hexokinase,
glucose-6- phosphatase, Fructose-1,6bi phosphatase were observed
in diabetic rats. After the Cinnamomum zeylanicum treatment
their activities reverted to near normal level.

Overall, it may be concluded that Cinnamomum zeylanicum

extract possesses hypoglycemic potential and has been shown to
afford significant protection against alloxan diabetes.

Histopathlogical studies in pancreas

Histopathological examination of pancreatic section of

control groups (fig) showed normal cellular architecture with
distinct β islet cells, sinusoidal spaces and central vein.

Pancreatic sections of the rat stained who hemtoxylin and

eosin (H&E) showed that alloxan caused disarrangement and
degeneration with severe necrotic changes of pancreatic islets,
especially in the centre of the islets. Nuclear changes, karyolysis,
disappearance of nucleus and in some places, residue of destroyed
cells were visible. Relative reduction of size and a number of
islets especially around the central vessel and severe reduction of
β-cells was clearly seen (fig)

Study of pancreas of treated diabetic group DG-1 & DG-2

Showed increased size of values and hypo chromic nucleus in
section stained with H&E. There was also a relative increase of
granulated and normal beta cells in the diabetic group when
compared with the diabetic group which consume 200mg/kg
weight Cinnamomum zeylanicum extract (fig) and Glibenclamide
which consume 600mg/kg body weight (fig).
Pancreas of the non-diabetic group which consume
200mg/kg body weight cinnamomum zeylanicum extract (fig
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