Antinephrolithiatic Potential of Macrotyloma uniflorum Aqueous Extract in Rats
VDN Srinivas*, G Sumalatha, PPV Jagannath, D Siva Kishore, P Madhu Kiran and M Jhansi Rani
Department of
Pharmacology, Hindu College of Pharmacy, Amaravathi
Road, Guntur, Andhra pradesh
ABSTRACT:
Urolithiasis is a major health problem with its high
morbidity, high cost of management and potential for end-stage renal disease.
The aim of our study is to cure and prevent the recurrent stone formation.
Herbal drugs claim many promising remedies in urolithiasis.
An attempt is made in this direction in the current work by using Macrotyloma uniflorum.
The experimental model selected for the present study is subcutaneous
administration of gentamicin (40mg/kg/day) for 8 days
and dietary supplementation with 5% ammonium oxalate starting at day 5 of the
treatment regimen to induce calcium oxalate urolithiasis.
On administration of aqueous extract of Macrotyloma uniflorum in
preventive and curative groups, moderate to few crystals are observed along the
mild appearance of edema and dilation in tubules and crystals are present
focally indicating the ability of aqueous extract of Macrotyloma uniflorum to
dissolve the pre-formed stones. The parameters monitored in the present study
are Kidney weight, calcium and oxalate deposition in kidney and the
histological studies support the calcium and oxalate deposition data in kidney.
Thus aqueous extract of Macrotyloma uniflorum showed significant antinephrolithiatic
activity.
KEYWORDS: Urolithiasis, Macrotyloma
uniflorum, calcium oxalate stones.
INTRODUCTION:
The incidence of urolithiasis is high in developing countries like and the
northern and north-western regions of
Urolithiasis is the third most
common disease of the urinary tract and it is affecting the human beings since
the earliest days. The recognition of different types of urinary calculi also
resulted in more varieties of medical treatment. Still recurrence rates
continue to be high with one of every two patients having another stone with in
5 years1. So, there is a need to develop new drugs to prevent the
recurrence of kidney stones. . In modern era, herbals are seen as
potential medicine for a variety of diseases often viewed to supercede the
pharmacological efficacy of allopathic drugs.
In most cases of the urolithiasis, the common component is either calcium
oxalate (75-90%) or magnesium ammonium phosphate to an extent of 5-10%2, 3.
In Indian indigenous system of medicine Macrotyloma uniflorum (Fabaceae)
is reported to be useful in the treatment of urinary stones. Although the plant
is claimed to be useful in the treatment of urinary stones, there is no record
of systematic pharmacological studies on the plant.
Figure-1
Figure-2:
PREVENTIVE CONTROL
PREVENTIVE
Figure-3:
CURATIVE CONTROL
CURATIVE
The present study was planned to assess the
efficacy of the aqueous extract of seeds of Macrotyloma uniflorum as antinephrolithiac
in albino rats. The animal model of nephrolithiasis
involving tubular dysfunction or injury by administration of gentamicin parenterally and 5%
ammonium oxalate mixed with standard rat pellet feed to induce calcium oxalate
stones.
MATERIALS
AND METHODS:
Source
of Plant Seeds:
The plant seeds were collected from Basavaiah Ayurvedic Pharmacy, Chintaluru,
Andhra Pradesh.
Preparation
of aqueous extract:
Shade dried powdered seed material was mixed
with water in 1:3 ratios for half an hour by heating over a water bath, kept
overnight and filtered. The filtrate was concentrated (120mg/ml) in a china
dish by heating over a water bath and was directly administered to animals.
PHARMACOLOGICAL
STUDIES:
Experimental
animals:
Male albino rats of Wistar
strain weighing between 150-200gm were used. The animals were fed with
commercial rat feed pellets (gold mohur) and were
given water Ad libitum. They were kept in
polypropylene cages in well-ventilated room under hygienic conditions.
Acute
toxicity studies:
Healthy adult albino rats were fasted
overnight with free access to drinking water. They were divided into five
groups each containing eight animals. Group-I animals were treated with distilled
water (2ml/kg/p.o) and Group-II to Group-IV animals received 1, 2, 4 and 8gm/kg
p.o of aqueous extract of Macrotyloma uniflorum respectively.
TABLE
-1: Effect of Aqueous Extract of Macrotyloma Uniflorum on Kidney Weights
|
S. No |
Group |
Kidney
weight gm/ 100gms of body weight |
|
I |
|
0.4± 0.013 |
|
II |
PREVENTIVE-CONTROL |
0.56±0.015* |
|
III |
PREVENTIVE |
0.33±0.016** |
|
IV |
CURATIVE-CONTROL |
0.5±0.016* |
|
V |
CURATIVE |
0.39±0.0127** |
*P<0.001 compared to normal
**P<0.001 compared to respective control
groups
TABLE -2:
Effect of Aqueous Extract of Macrotyloma Uniflorum on Deposition of Calcium and Oxalate in
Kidney
|
S. No. |
Group |
Deposition
in kidney weight mg / gm of weight tissue |
|
|
|
|
Calcium |
Oxalate |
|
I |
|
0.52±0.07 |
2.42±0.2 |
|
II |
PREVENTIVE – CONTROL |
2.5±0.3* |
10.09±1.21* |
|
III |
PREVENTIVE |
0.31±0.03** |
5.65±0.36** |
|
IV |
CURATIVE-CONTROL |
2.45±0.2* |
13.4±0.63* |
|
V |
CURATIVE |
0.96±0.1* |
8.69±0.23** |
*P<0.001 compared to normal
**P<0.001 compared to respective control
groups
The animals were observed continuously for 2
hours, and then intermittently and at the end of 48 hours, the number of deaths
was noted to calculate LD504.
Gross behavioral changes:
The animals were observed for behavioral,
neurological and autonomic profiles during acute toxicity studies5
Antinephrolithiatic study-
Calculi Producing Diet (CPD) MODEL:
To study the effect of aqueous extract of Macrotyloma uniflorum on various
parameters of stone formation such as
1. Effect of extract on kidney weight. (Table
1)
2. Deposition of calcium and oxalate in
kidney. (Table 2)
3. Histological studies of kidney. (Fig 1-3)
In the nephrolithiatic
study gentamicin treatment predispose to stone
formation by inducing some damage i.e. focal necrosis, loss of border membranes
and the occurrence of membrane debris in the tubule lamina. So, membrane debris
formed due to renal damage can act as nucleation sites for calcium oxalate
crystallization. This pattern of calculi deposition in renal medulla region is
similar to human urolithiasis. Gentamicin
has been shown to inhibit calcium reabsorption in
proximal tubule6. Kidneys of nephrolithiatic
rats showed deposition of calcium oxalate crystals in renal calculi and at the
papillary tips. Many were located sub epithelial often anchored to the basement
membrane. Hyperoxaluria and calcium oxalate
deposition was induced by gentamicin and calculi
producing diet. The standard rat pellet feed was powdered and mixed with
ammonium oxalate (5%) and again made into pellets. This feed is called as
calculi producing diet (CPD) and it is fed to animals to induce hyperoxaluria and calcium oxalate calculi. The animals were
divided into five groups of ten animals each and were put on treatment
schedules as shown in Fig.4.
Administration of gentamicin
(40mg/kg/s.c) for first eight days induced renal tubular damage and hemorrhages
in the kidneys of experimental animals. Co-administration of calculi producing
diet (5%ammonium oxalate with standard rat feed) caused an increase in the
deposition of calcium and oxalate in the kidney and urinary excretion of calcium
and oxalate in both the control groups when compared to normal animals.
Effect
of Extract on Kidney weight:
A significant increase in kidney weight was
noted in the preventive control and curative-control groups when compared to
normal animals (p<0.001) (Table 1).
A significant decrease in kidney weight was
observed in the preventive and curative groups treated with aqueous extract of Macrotyloma uniflorum when
compared to their respective control groups (p<0.001).
Deposition
of calcium and oxalate in kidney:
A significant increase (p<0.001) in the
deposition of calcium and oxalate was observed in both preventive-control and
curative-control groups when compared to normal animals.
Figure 4:
Treatment Schedules For Antinephrolithiatic Study
After administration of aqueous extract of Macrotyloma uniflorum for 15 days
in the preventive group, a significant (p<0.001) decrease in calcium and
oxalate deposits were observed when compared to the preventive-control group.
These results indicate the efficacy of aqueous extract of Macrotyloma
uniflorum in preventing the calcium oxalate type of
stone formation in the kidney.
In the curative group, administration of
aqueous extract of Macrotyloma uniflorum
for 15 days caused a significant decrease (p<0.001) in calcium and oxalate
deposition when compared to the curative-control group. This evident the
significant effect of the aqueous extract of Macrotyloma
uniflorum in dissolving the pre-formed calcium
oxalate type of stones.
The calculi producing diet (CPD) has some
advantages like:
·
Rapid
crystal and stone deposition with in 14 days.
·
The
pattern of stone deposition resembles that of human stone disease as majority
of the crystals occur in the papillae.
·
Renal
function remains relatively normal.
Statistical Analysis:
The results are expressed as mean ± SEM and
were statistically analyzed by Students t- test7.
RESULTS
AND DISCUSSION:
Administration of aqueous extract of Macrotyloma uniflorum
significantly reduced both calcium and oxalate deposition in the kidneys of
both preventive and curative groups when compared to their respective controls.
This indicates that aqueous extract of Macrotyloma uniflorum has beneficial effect in preventing calculi
formation.
On histopathological
examination both the preventive and curative control groups showed presence of
calcium oxalate crystals in Lumina of tubules accompanied by edema and cast
formation extensive intertubular hemorrhages and
congestion of blood vessels. These histological observations support the
presence and growth of renal calculi in renal medulla region as observed in
human urolithiasis. On administration of aqueous
extract of Macrotyloma uniflorum in
preventive and curative groups moderate to few crystals are observed along the
mild appearance of edema and dilation in tubules and crystals are present
focally indicating the ability of aqueous extract of Macrotyloma uniflorum to dissolve the pre-formed
stones. These histological studies support the calcium and oxalate deposition
data in kidney. Thus aqueous extract of Macrotyloma uniflorum showed significant
antinephrolithiatic activity.
CONCLUSION:
The present study has shown that formed
urinary stones could be dissolved with aqueous extract Macrotyloma uniflorum and without the aid of surgical intervention. The
recurrence of stones could also be prevented to a great extent. While it may
not be possible straightway to altogether dispense with surgical and other
invasive procedures, the use of Antinephrolithiatic
drugs is definitely going to revolutionize the treatment of urinary stones.
ACKNOWLEDGEMENTS:
The authors are Thankful to Hindu College of
Pharmacy for providing necessary laboratory facilities to carry out present
research work.
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Received on 07.07.2009
Accepted on 10.08.2009
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all right reserved
Research Journal of Pharmacognosy and Phytochemistry. 2(1): Jan.-Feb. 2010, 75-78