Phytochemical Screening and Antihyperglycemia

activity of Aqueous Bougainvillea spectabilis extract as

a Potential Source of Anti-hyperglycemic Agents: A

Clinical Trial

AN SIP PROPOSAL

SUBMITTED TO HIMAMAYLAN NATIONAL HIGHSCHOOL

LIFE SCIENCE (TEAM CATEGORY)

PROPONENTS:

ZALDY MAR L. LAVADA JR.

GAZYL V. TALORONG

KANEESHA KHYNS P. GAYARES

AUGUST 4 , 2017

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
 TABLE OF CONTENTS

PART PAGE NUMBER

I.INTRODUCTION

I.1 Background of the Study 3

I.2 Objectives of the Study 4

I.3 Significance of the Study 4-5

I.4 Scopes and Limitations 5

I.5 Hypotheses 5-6

II. Methodology

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
 I. Introduction

I.1 Background of the study

The quantity of the trials men are facing right now is limitless.First on list are

the diseases that are being caused by a sedentary type of lifestyle .Take diabetes for

an instance; it conjures an abnormally high amount of sugar present in the human

blood.However , humanity handled this disease by allowing synthetic insulin to

circulate in the human body but its availability is hurdled by its high price.

Several alternatives for insulin were made like the miracle fruit and the acacia

species of trees.But another one is as effective as the mentioned plants are.No

expensive costs and it can be found almost everywhere - in our yards or beside

highway roads because of its beautiful flowers.The Bougainvillea spectabilis,

commonly known as the Bougainvillea plant is also one of the 30 000 - 70 000 [1]

(Fabricant, 2001) plant species that are believed to have healing effects yet people

have no clue about it.

This project investigates the glucose-reducing effects of the aqueous extract of

Bougainvillea flowers and leaves to prove the potential medicinal use of the plant.

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
I.2 Objectives of the Study

 To identify the glucose reducing properties of Bougainvillea spectabilis

aqueous extract.

 To determine which between the leaves and the flowers is more effective.

I.3 Significance of the Study

These critical times, where various diseases are prominent all throughout the

globe; plants became the vital cog of most scientists .They are used as effective

alternatives for synthetic drugs such as insulin.

A single syringe with insulin costs ₱200 . And most people, especially those

under the poverty line will have difficulties in purchasing a medicine with such a

high price.

To this effect, this project determines the potential of Bougainvillea spectabilis

as a cheaper and a more available alternative for synthetic drugs in lowering the

blood sugar. And therefore bring great opportunities to the poorer population to

have a natural alternative that can help lower the blood sugar of persons with

hyperglycemia. It could also open doors leading to a considerable amount of

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
income because its availability is unrivalled by any other synthetic and natural

remedies in our tropical country.

I.4 Scopes and Limitations

This study was conducted to determine the capability of the bougainvillea

spectabilis in reducing the blood sugar levels of hyperglycemic mice.This also

identifies the phytochemical constituents of Bougainvillea spectabilis namely D-

pinitol. Extracted leaves and flowers were used to determine the Anti-

hyperglycemia effects on a group of wistar mice obtained from Himamaylan City.

The invetigation was done at Riverside College Incorporated Laboratory

at Bacolod City , Negros Occidental Philippines on the month of August 2017.

I.5 Hypotheses

Alternative Hypotheses:

1. The Bougainvillea plant will be effective in neutralizing excessive sugar in

the blood.

2. The Bougainvillea leaves will deal the most significant amount of decrease

in the glucose – level.

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
Null hypothesis :

1. There will be no significant difference in the sugar levels of the

hyperglycemic mice that were injected with bougainvillea extract and those

who are left untreated.

II. Methodology

II.1 Procedure

II.1.1 Collection of Test Subjects

Month-old wistar mice were bought in Bacolod City, Negros

Occidental Philippines. They were previously fed with commercial food

and was identified with normal blood sugar.

II.1.2 Collection Of Plant Materials

Bougainvillea spectabilis plants were obtained from Silos

Subdivision, Brgy.Talaban , Himamaylan City Negros Occidental.

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
 I. Review of Related Literature

Review Related Literature

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
• Hypoglycemic / Stem Bark:Study evaluated the hypoglycemic potential of B.

spectabilis stem bark extract in albino rats. Results showed significant anti-

hyperglycemic effect by the stem bark, 22.2% more potent than oral hypoglycemic

drug, glibenclamide 0.2 mg/kg.

• Pinitol / Insulin-like effect: Pinitol, an active principle of the traditional

antidiabetic plant B. spectabilis, is claimed to exert insulin-like effects. The study

supported the view that D-pinnitol (3-O-methyl-chiroinositol) may exert an

insulin-like effect to improve glycemic control in hypoinsulinemic STZ-diabetic

mice. D- pinitol may act via a post-receptor pathway of insulin action affecting

glucose uptake.

• Antibacterial / Leaves: Study on various solvent extracts of Bougainvillea

spectabilis leaves showed maximum inhibitory effect on tested bacteria (S aureus,

B subtilis, S faecalis, Micrococcus luteus, E coli, P aeruginosa, S typhii, K

pneumonia, P vulgaris, S marcescens, S flexneri. Study of B. spectabilis and B.

variegata ethanolic and methanolic extracts of leaves showed significant

antimicrobial activity.

Anti-diabetic Properties of the Aqueous Leaf Extract of Bougainvillea glabra

(Glory of the Garden) on Alloxan-Induced Diabetic Rats

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
Abstract: The antidiabetic and antilipidemic effects of Bougainvillea glabra was

investigated in this study using 25 male wistar rats. The rats were divided into 5

groups comprising of five animals each. These groups include a normal control

(administered saline), an extract control (administered 100 mg/kg of extract) and a

diabetic control (untreated group). The remaining two groups were administered

100mg/kg and 400 mg/kg of the extract respectively. The study lasted for three

weeks although blood samples were obtained from the rat tails after every week.

The results show that the extract significantly (p< 0.05) reduced the

hyperglycaemia from 12±0.40 mmol/L (Diabetic Control) to 4.04±0.03 mmol/L

(400 mg/kg group). Likewise, the extract significantly reduced the Total

Cholesterol (TC), Triglyceride (TG) and Low-Density Lipoprotein Cholesterol

(LDLCholesterol), while increasing the High-Density Lipoprotein Cholesterol

(HDL-C). In conclusion, the observations from this study show that Bougainvillea

glabra has antidiabetic effect and beneficial effects on blood lipid profile, thus

justifying the use of the plant by traditional medicine practitioners for the treatment

of diabetes mellitus

Isolation of Antidiabetic Principle from Bougainvillea spectabilis Willd

(Nyctaginaceae) Stem Bark

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
Abstract:Purpose: To isolate and identify the constituents of Bougainvillea

spectabilis Willd (Nyctaginaceae) stem bark.

Methods: The methanol extract of Bougainvillea spectabilis stem bark powder

was suspended in water and extracted with dichloromethane (CH2Cl2), ethyl

acetate (EtOAc), and butanol (BuOH) successively. The ethyl acetate fraction was

loaded in a column packed with silica gel and eluted with a gradient of chloroform

(CHCl3): methanol (MeOH), and water yielded five fractions (A - E). Chemical

constituents were isolated by repeated column chromatography of these fractions.

Results: Column chromatography of fractions B and C afforded four compounds

identified as pinitol, β-sitosterol, quercetin and quercetin-3-O-α-L-

rhamnopyranoside. For the first time, pinitol, β-sitosterol, quercetin and quercetin-

3-O-α-L-rhamnopyranoside were isolated from the stem bark of B.

spectabilis Willd.

Hypoglycemic activity of Bougainvillea spectabilis stem bark in normal and

alloxan-induced diabetic rats

Abstract

Bougainvillea spectabilis (B. spectabilis) is one of the main constituent of various

herbal formulations available for diabetes. The aim of present study was to screen

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
hypoglycemic potential of B. spectabilis stem bark extracts in albno rats (Wistar

strain).

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
Antihyperlipidemic effect of D-pinitol on streptozotocin-induced diabetic

wistar rats

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
Abstract; D-pinitol (3-O-methyl-chiroinositol), an active principle of the

traditional antidiabetic plant, Bougainvillea spectabilis, is claimed to exert

insulin-like effects. This study was undertaken to evaluate the effect of D-

pinitol on lipids and lipoproteins in streptozotocin (STZ)-induced diabetic

Wistar rats. Rats were made type II diabetic by single intraperitoneal

injection of STZ at a dose of 40 mg/kg body weight. STZ-induced diabetic rats

showed significant (p < 0.05) increase in the levels of blood glucose and total

cholesterol, triglycerides, free fatty acids, and phospholipids in serum, liver,

kidney, heart, and brain. The levels of low-density lipoprotein (LDL) and very

low-density lipoprotein (VLDL) cholesterol were significantly increased, and

the level of high-density lipoprotein (HDL) cholesterol was significantly

decreased in diabetic rats Oral administration of D-pinitol to STZ-induced

diabetic rats showed significant (p < 0.05) decrease in the levels of blood

glucose and total cholesterol, triglycerides, free fatty acids, and phospholipids

in serum, liver, kidney, heart, and brain. The D-pinitol also lowered

significantly (p < 0.05) LDL and VLDL cholesterol levels and increased

significantly (p < 0.05) HDL cholesterol levels in the serum of diabetic rats.

Thus, the present study clearly showed the antihyperlipidemic effect of D-

pinitol in STZ-induced type II diabetic rats

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
HYPOGLYCEMIC POTENTIAL OF BOUGAINVILLEA SPECTABILIS

ROOT BARK IN NORMAL AND ALLOXANINDUCED DIABETIC RATS

Introduction: Diabetes is a lifelong (chronic) metabolic disorder which is caused by

deficiency of insulin, or ineffectiveness of insulin produced . Type-1 diabetes may

be due to deficiency of insulin secretion-due to destruction of pancreatic β cells

and the type-2 diabetes may be due to insulin resistance- a defect in the tissue

response to insulin and impaired insulin release- slow basal secretion of insulin

after meal . Premature illness and death due to diabetes has reached epidemic

proportions worldwide . World Health Organization estimates that developing

countries will be most affected by this epidemic in the present 21st century The

global figure of patients with

Bougainvillea spectabilis Exhibits Antihyperglycemic and Antioxidant

Activities in Experimental Diabetes

Abstract

The study investigates the effects of aqueous extract of Bougainvillea

spectabilis leaves on blood glucose, glycosylated hemoglobin, lipid profile,

oxidative stress, and on DNA damage, if any, as well as on liver and kidney

functions in streptozotocin-induced diabetes in Wistar rats. Daily administration

of the aqueous extract of B spectabilis leaves for 28 days resulted in significant

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
 reduction in hyperglycemia and hyperlipidemia as evident from restoration of

relevant biochemical markers following extract administration. The extract also

exhibited significant antioxidant activity as evidenced from the enzymatic and

nonenzymatic responses and DNA damage markers. The extract restored kidney

and liver functions to normal and proved to be nontoxic. A marked improvement

in the histological changes of tissues was also observed. The present study

documented antihyperglycemic, antihyperlipidemic, and antioxidative potentials

of the aqueous extract of B spectabilis leaves without any toxicity in

streptozotocin-treated Wistar rats.

An overview on antidiabetic medicinal plants having insulin mimetic property

Abstract

Diabetes mellitus is one of the common metabolic disorders acquiring around 2.8%

of the world's population and is anticipated to cross 5.4% by the year 2025. Since

long back herbal medicines have been the highly esteemed source of medicine

therefore, they have become a growing part of modern, high-tech medicine. In

view of the above aspects the present review provides profiles of plants (65

species) with hypoglycaemic properties, available through literature source from

various database with proper categorization according to the parts used, mode of

reduction in blood glucose (insulinomimetic or insulin secretagogues activity) and

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
active phytoconstituents having insulin mimetics activity. From the review it was

suggested that, plant showing hypoglycemic potential mainly belongs to the family

Leguminoseae, Lamiaceae, Liliaceae, Cucurbitaceae, Asteraceae, Moraceae,

Rosaceae and Araliaceae. The most active plants are Allium sativum, Gymnema

sylvestre, Citrullus colocynthis, Trigonella foenum greacum, Momordica

charantia and Ficus bengalensis. The review describes some new bioactive drugs

and isolated compounds from plants such as roseoside, epigallocatechin gallate,

beta-pyrazol-1-ylalanine, cinchonain Ib, leucocyandin 3-O-beta-d-galactosyl

cellobioside, leucopelargonidin-3- O-alpha-L rhamnoside, glycyrrhetinic acid,

dehydrotrametenolic acid, strictinin, isostrictinin, pedunculagin, epicatechin and

christinin-A showing significant insulinomimetic and antidiabetic activity with

more efficacy than conventional hypoglycaemic agents. Thus, from the review

majorly, the antidiabetic activity of medicinal plants is attributed to the presence of

polyphenols, flavonoids, terpenoids, coumarins and other constituents which show

reduction in blood glucose levels. The review also discusses the management

aspect of diabetes mellitus using these plants and their active principles.

Anti-diabetic and hypolipidemic effects of the aqueous leaf extract of

Bougainvillea species

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
Bougainvillea (with the common name Glory of the Garden), originated from

South America and it is a popular plant in Southern California, Florida, the

Caribbean and other areas with tropical and warm climates. It is a thorny woody

plant with flowers ranging from pink, purple, red, orange, yellow colors and

especially white. Among the Bougainvillea’s varieties include B. glabra,

Bougainvillea spectabilis, and Bougainvillea harrisi.

It is also observed that B. glabra and B. spectabilis have some anti-lipidemic

effects. high-density lipoproteincholesterol levels increased in the diabetic mice

groups who were administered the Bougainvillea extracts. Major complications of

diabetes include aberrant lipid metabolism and vascular wall function. Since

alterations in serum lipid profiles are known in diabetes which are likely to

increase the risk of coronary heart disease, a reduction in serum lipids, particularly

total cholesterol, low-density lipoproteincholesterol and triglycerides levels should

be considered as beneficial in long-term prognosis of diabetic patients. B. glabra

extract will thus have a potential therapeutic value in combating multifactorial

atherosclerotic disorders, which are parts of the major complications of DM.

Reducing the risk of atherosclerosis will thus lead to the development of effective

and better management of hyperlipidemia.

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
PHYTOCHEMICAL AND IN-VITRO ANTHELMINTIC ACTIVITY OF

HYDRO ALCOHOLIC EXTRACT OF BOUGAINVILLEA GLABRA

ABSTRACT

Bougainvillea glabra, a flowering plant generally used as ornamental plant and

seen mostly in areas with warm climate. Apart from its use as ornamental plant, the

leaf of B.glabra is reported to have various medicinal values. This current study

deals with preliminary phytochemical evaluation and determination of anthelmintic

activity of hydro alcoholic leaf extract of Bougainvillea glabra. The study includes

preparation of extract by solvent extraction method with soxhlet apparatus using

hydro alcohol and petroleum ether as solvents. Then both extracts are evaporated

separately at ambient temperature to concentrate it. Preliminary qualitative

chemical test done for both extracts show the presence of alkaloids, glycosides,

carbohydrates, anthraquinone, flavanoids, terpenoids, saponins, steroids, proteins,

fixed oils, fats and tannins. The in-vitro anthelmintic activity of extracts performed

with the Indian earth worm shows better results when compared with

Metronidazole as the standard drug.

Phytochemical Screening and Antibacterial Acitivity of Bougainvillea

Glabra Plant Extract as Potential Sources of Antibacterial and Resistance-

modifying Agents
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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.
 The crude methanol extract of different parts of Bougainvillea glabra were

experimented and examined for its Phytochemical Constituents and

Antibacterial Activity Index with four Commercial Antibiotics against 4

bacterial strains. Phytochemical screening revealed the presence of alkaloids,

saponins, tannins, reducing sugar and cardiac glycosides in the leaves of

Bougainvillea glabra. Antrhaquinones, flavonoids and terpenoids were absent in

the leaves of Bougainvillea glabra. Different parts of Bougainvillea glabra have

shown antibacterial potentials when used on two gram-positive bacteria, the

Bacillus Subtilis and Staphylococcus aureus; and two gram-negative bacteria,

the Escherichia coli and Pseudomonas aeruginosa except for flowers against two

gram-positive bacteria, the Bacillus Subtilis and Staphylococcus aureus. Results

show that this plant is an interesting source of antimicrobial agents, potential

source of nutraceuticals, anti-inflammatory, antiallergen

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Lavada, Zaldy Mar L., Gayares,Kaneesha P., Talorong, Gazyl V.