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com 110
International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701
_________________________________________Research Paper
Protective Effect of Aqueous Extract of Uraria Picta on
Acetaminophen I nduced Nephrotoxicity in Rats
Kale R. H.
1*
, Halde U. K.
2
and Biyani K. R.
1

1
Anuradha College of Pharmacy, Chikhli, Maharashtra, India.
2
Drug Testing Laboratory, Government Ayurvedic and Unani Pharmacy, Nanded,
Maharashtra, India.
__________________________________________________________________________________
ABSTRACT
Nephrotoxicity can be produced by the excess use of NSAID (Non steroidal anti-inflammatory drugs) like
Acetaminophen, it-induced liver necrosis and renal insufficiency occurs in approximately 12%of patients with
acetaminophen overdose. Uraria picta is one of the important constitutents among the ten herb formulation
called Dashmula. Its content flavonoid are known to exhibit a range of biological activities like anti-
inflammatory, anti-thrombotic, hepatoprotection properties due to their free radical scavenging ability.
Healthy wistar albino rats (180-240 g) divided into four groups each containing six rats. Group I was taken as a
normal, group II treated with 2.5 g/kg Acetaminophen and group III and group IV treated with Aqueous extract
of Uraria picta plus acetaminophen (250 and 500 mg/kg dose) administered for seven days. Treatment with
the aqueous extract of Uraria picta (250 and 500 mg/kg/body wt.) containing polyphenolic compound and
carbohydrates might be significantly reduces the elevated levels of urine urea and BUN levels and also
elevated levels of Serum Creatinine and Urine creatinine compared to acetaminophen group. The activity
elicited by the extract might be due to its ability to activate antioxidant enzymes. The findings suggest the
potential use of the aqueous extract of Uraria picta as a novel therapeutically useful nephroprotective agent.

Key Words: Acetaminophen, Histopathology, Nephrotoxicity, Uraria picta.

INTRODUCTION

Nephrotoxicity is a poisonous effect of some
substances, both toxic chemicals and medication,
on the kidneys. There are various forms of toxicity,
Nephrotoxicity should not be confused with the
fact that some medications have a predominantly
renal excretion and need their dose adjusted for the
decreased renal function. Mainly nephrotoxicity
can be produced by the excess use of NSAID (Non
steroidal anti-inflammatory drugs) like
Acetaminophen, it-induced liver necrosis and renal
insufficiency occurs in approximately 12% of
patients with acetaminophen overdose. Heavier
acetaminophen use was associated with an
increased risk of end-stage renal disease in a dose
dependent fashion
1
.
At therapeutic doses, acetaminophen is metabolized
via glucuronidation and sulfuration reactions
occurring primarily in the liver, and results in
water-soluble metabolites that are excreted via the
kidney. As a result of the metabolic conversion of
acetaminophen by the microsomal P-450 enzyme
system, a highly reactive intermediate, N-acetyl-
pbenzoquinone imine (NAPQI) is produced.
NAPQI directly reacts with glutathione (GSH) and
at overdoses of acetaminophen, the depletion of
cellular GSH occurs. This allows NAPQI to bind to
cellular proteins and initiate lipid peroxidation,
leading to renal injury previous studies
demonstrated that acute APAP overdose increased
lipid peroxidation, endoplasmic reticulum stress.
Paracetamol (also known as acetaminophen)
poisoning is due to metabolic activation of APAP
by renal P450 mixed-function oxidases, similar
mechanism was proposed for the
nephrotoxicity
2,3,4
.
Uraria picta is one of the important constitutents
among the ten herb formulation called Dashmula, a
well established Ayurvedic drug of the Indian
system of medicines for treating general fatigue,
oral sores and several gynaecological disorders
5
.
Its content flavonoid are known to exhibit a range
of biological activities like anti-inflammatory, anti-
thrombotic, hepatoprotection properties due to
their free radical scavenging ability. These
observations support the mechanism of
nephrotoxicity induced by acetaminophen in
animals is partially related to the depletion of renal
antioxidant system
6
.





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International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701
METHODS AND MATERIALS

Plant material
Whole plant of Uraria picta Desv. was collected
fromShri. Ganesh Nursery Jeur Purender Pune and
also procure authentified aqueous extract of Uraria
picta Desv. from Amsar Pvt. Ltd. Indore (MP). The
whole plant was authentified in Botany department
of the Shivaji Science College Chikhli Dist.
Buldana (MS). India.

Animals
Male albino Wistar rat 6-8 weeks of age and
weighing between (180240) gm were selected
fromAnuradha College of pharmacy, Chikhli, Dist-
Buldana (MS) India. They were housed in
polypropylene cages under the standard conditions
of temperature, pressure and humidity. (12 hrs light
and dark cycles, at 25 + 3C and 35-60%
humidity).The animals were maintained under
standard condition as per CPCSEA guideline.
Animals were fed with standard normal pellet diet
and water ad libitum during the course of the
experiment, according to the guidelines of
Institutional Animal Ethical Committee (IAEC) of
Anuradha College of pharmacy, Chikhli, Dist-
Buldana (MS) India, registered under CPCSEA,
India (Registration no. abc/751/03/CPCSEA).

EXPERIMENTAL DESIGN

Healthy wistar albino rats (180-240 g) divided into
four groups each containing six rats. Group I was
taken as a normal treated with normal saline, group
II treated with 2.5 g/kg Acetaminophen was kept as
a control (20% suspension in saline stabilized by
0.2% gum) by means of a stomach tube, group III
and group IV treated with Aqueous extract of
Uraria picta plus acetaminophen (250 and 500
mg/kg dose) administered for seven days. 48 hours
after the acetaminophen injection animals were
sacrificed using ether anesthesia blood was
collected fromeach animal. Serumwas used for the
determination of ureaand creatinine by Span
diagnostic kit (Span Diagnostics Ltd., India. The
elevation of urea and creatinine level in the serum
was taken as the index of nephrotoxicity.

EVALUATION OF RENAL FUNCTION

Blood and urine urea
Urea concentration in the blood and urine were
estimated by enzymatic method using Urease
enzyme kit (DAM kit) by colorimetric end point
method. Absorbance was read using
photometrically at wavelength 540 nm.

Serum and urine creatinine
Creatinine level in the serum and urine were
estimated by the alkaline picrate method using
creatinine kit method. (Jaffes modified kinetic
method) and the absorbance was read at 520 nm
(Span diagnostics Ltd).

Histopathological Study
The two animals from each group were sacrificed
on the day of blood withdrawal and kidneys were
isolated. Tissue samples were immersed in10%
formalin for at least 24 hr to fix the tissue, the
tissue was then embedded in paraffin wax,
sectioned and stained with haematoxylin and eosin.
The sections were then viewed under the light
microscope for Histopathological changes.

Statistical analysis
The data were presented as Mean SD and the
statistical analysis by one way ANOVA followed
by Dunnetts multiple comparison tests.

RESULTS AND DISCUSSION

Table 1 and 2 shows, urine urea and BUN levels
were significantly elevated in the acetaminophen
treated group compared to the normal group
indicating the induction of severe nephrotoxicity.
Treatment with the aqueous extract of Uraria picta
(250 and 500 mg/kg/body wt.) containing
polyphenolic compound and carbohydrates might
be significantly reduces the elevated levels of urine
urea and BUN levels and also elevated levels of
SerumCreatinine and Urine creatinine compared to
acetaminophen group (P<0.0001).
Figure 1(A) indicate normal renal tissue without
any disfunction. Light microscopy of renal tissue
(Fig. 1(B)) in acetaminophen treated rats. Sever
tubular necrosis, tubular degeneration and cellular
infiltration are observed. Moderate necrosis,
tubular degeneration, and cellular infiltration are
shown in Fig. 1(C) on administration of 250 mg/kg
aqueous exract.while Light microscopy of renal
tissue in Fig. 1(D), normal architecture of renal
tissue is observed on administration of 500 mg/kg
aqueous extract.
Many metabolic disorders including serum
electrolyte, urea and creatinine dearrangements is
found due to over dose of acetaminophen.
Increased concentration of serum urea and
creatinine are considered for investigating drug
induced nephrotoxicity in animals and man
7
. Blood
urea nitrogen is found in the liver protein that is
derived fromdiet or tissue sources and is normally
excreted in the urine. In renal disease, the serum
urea accumulates because the rate of serum urea
production exceeds the rate of clearance
8
. Elevation
of urea and creatinine levels in the serum was 96
taken as the index of nephrotoxicity
7, 9
. In this
study, acetaminophen Induced nephrotoxicity
showed a significant (P<0.001) increase in the
serum urea and creatinine concentrations in the
Group II (acetaminophen induced) rat when

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International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701
compared to the normal group (Group I).
Moreover, oral administration of aqueous extract of
Uraria picta significantly (P<0.01) decreased in
group III and IV when compared to the Group II.
However the level of uric acid is significantly
increased (P<0.001) in the Group II rats when
compared to Group I. Creatinine, on the other
hand, is mostly derived from endogenous sources
by tissue creatinine breakdown
10
. Thus serum urea
concentration is often considered a more reliable
renal function predictor than serumcreatinine. In
the present study, administration of nephrotoxic
doses of acetaminophen to rats resulted in
development of oxidative stress damage in renal
tissues.



Table 1: Effect of aqueous extract of Uraria picta on urine urea and Blood Urea Nitrogen level
Parameters
Normal
(Distilled water)
Toxicant
Control
(Acetaminophen)
Aqu. Extract
250mg/kg + Acetaminophen
Aqu. Extract
500mg/kg + Acetaminophen
Blood Urea
mGs/dL
12.321.03 26.590.97*** 24.820.57** 19.700.35**
Urine urea
(Gms/24hrs)
22.300.48 48.380.45*** 42.530.38** 38.130.28**
Values are expressed as themeanSEM (n=6) control groups was compared with treated groups.
Experimental groups compared with treated Groups. *p<0.05, **p<0.01, ***p<0.001.




Table 2: Effect of aqueous extract of Uraria picta on
Serum Creatinine and Urine creatinine Level
Parameters
Normal
(Distilled water)
Toxicant
Control
(Acetaminophen)
Aqu. Extract
250mg/kg
+ Acetaminophen
Aqu. Extract
500mg/kg
+ Acetaminophen
Serumcreatinine
(mg/100ml)
0.8360.26 2.360.142*** 1.410.10** 0.970.09***
Urine creatinine
(g/24 hrs)
2.060.097 3.090.036*** 2.850.038
ns
2.610.039**
Values are expressed as themeanSEM (n=6) control groups was compared with treated groups.
Experimental groups compared with treated Groups. *p<0.05, **p<0.01, ***p<0.001, ns=non significant




Fig. 1: Kidney section A (Normal), B) Acetaminophen, c) Aqueous extract (250 mg/kg)
and d) Aqueous extract (500 mg/kg)
Black arrow tubular degeneration and necrosis, Red arrow Congestion,
Blue arrow - Leucocytic infiltration, Orange arrow Glomerulopathy.



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International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701


Fig. 2: Uraria picta Plant
CONCLUSION

The Protection offered by the extract could have
been due to the presence of polyphenolic and
carbohydrates compounds. The activity elicited by
the extract might be due to its ability to activate
antioxidant enzymes. The findings suggest the
potential use of the aqueous extract of Uraria picta
as a novel therapeutically useful nephroprotective
agent. Therefore, further studies to elucidate their
mechanisms of action should be conducted to aid
the discovery of new therapeutic agents for the
treatment of renal diseases.

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