Renal Protectant Activity of
Cochlearia armoracia in 5/6-Nephrectomized Rat Model
Shanish Antony A.*, Jayasankar K., Partha Deb Roy, Pankaj
N., Dhamodaran P., Duraiswamy B. and Elango K.
Department of
Pharmacology, JSS College of Pharmacy, Ooty-643001, Tamilnadu, India
ABSTRACT:
We investigated the effect of aqueous and ethanolic
extract of Cochlearia armoracia at a
dose of 200 and 400mg/kg in 5/6 nephrectomized rats for its nephroprotective
activity. Blood samples were collected for the measurement of serum creatinine,
urea, blood urea nitrogen, uric acid. Urine samples were collected for the
measurement of urine volume, sodium and potassium excretion levels. In
nepherctomized rat the blood parameters were significantly increased and urine
parameters were significantly altered to disease condition. Both the extracts
at a dose level of 400mg/kg showed significant (p<0.01) decrease in serum
creatinine, urea, blood urea nitrogen, uric acid and increases urinary volume
and sodium and potassium excretions. The present study revealed that the oral
administration of aqueous and ethanolic extracts at 400mg/kg dose exhibit a
significant nephroprotective activity in 5/6 nephrectomized rat model.
KEYWORDS: Cochlearia armoracia; Nephrectomized and Nephroprotective
1. INTRODUCTION
The roots of Cochlearia armoracia Linn belongs to
family Cruciferae, is widely growing plant mainly in Western Ghats of Nilgiri
hills and was universally used as a medicine during middle age and as a
condiment in Denmark and Germany (Grieve, 1995; Bown,
1996).This root was included in Materia
Medica of London Pharmacopoeia of eighteen centuries. This
plant is available in few parts of Nilgiris and widely used as a traditional
medicine. The main phytoconstituents that are present are mainly glycoside and
coumarins (Hansen, 1974).
The roots of this plant is claimed to be used as a diuretic (Chopra et al.,
1992) and also
used to treat kidney disorders like kidney stones and edema (Goldberg et al., 1998). But scientifically the diuretic
activity has not reported and also the effect of Cochlearia armoracia in
chronic renal failure model has not yet established. In the present communication we report the renal protectant activity of
the root of Cochlearia armoracia in rat model of chronic renal failure.
2. METERIALS AND METHODS:
2.1. Collection of plant material:
The
roots of Cochlearia armoracia were collected during
June 2009, from the local areas of Nilgiris. The roots were identified and
authenticated by Botanist, Government Botanical Garden, Udagamandalam. The
voucher specimen was deposited in the herbarium of the department of Pharmacognosy,
JSS College of Pharmacy, Ooty.
Table
1: Effect of the aqueous and the ethanolic extracts on urinary and metabolites
excretion in nephrectomized rat after 15 days of treatment.
|
Groups |
Treatment |
Urinary
excretion ml/day |
Sodium ion
concentration (meq/day) |
Potassium ion
concentration (meq/day) |
|
1 |
Control |
7.67 ± 1.08 |
24.38 ± 2.12 |
30.52±3.01 |
|
2 |
Aqueous extract
200mg/kg |
10.45 ± 1.98** |
34.30 ± 0.81** |
41.56±2.14** |
|
3 |
Aqueous extract
400mg/kg |
14.95 ± 2.57** |
38.72 ± 1.56** |
48.83±1.65** |
|
4 |
Ethanolic extract
200mg/kg |
9.81 ± 1.28* |
32.81 ± 1.24** |
40.09±1.81** |
|
5 |
Ethanolic extract
400mg/kg |
12.91 ± 2.02** |
36.09 ± 2.57** |
46.92±1.63** |
Value represents mean ± SEM of six animals in each
group, Significant difference from the control, *P<0.05, **P<0.01.
Table 2: Effect of the aqueous and the ethanolic
extracts on creatinine, urea, uric acid and blood urea nitrogen in
nephrectomized rats after 15 days of treatment.
Groups
|
Treatment |
Creatinine (mg/dl) |
Urea (mg/dl) |
Blood
urea nitrogen (mg/dl) |
Uric acid (mg/dl) |
|
1 |
Control |
16.18 ± 0.47 |
170.8 ± 21.94 |
79.79 ± 10.24 |
11.23 ± 2.83 |
|
2 |
Aqueous extract
200mg/kg |
2.88 ± 0.19** |
26.08 ± 2.41** |
12.17 ±1.12** |
4.57 ± 1.52** |
|
3 |
Aqueous extract
400mg/kg |
1.03 ± 1.13** |
25.56 ± 1.13** |
11.93 ±0.52** |
2.59 ± 3.81** |
|
4 |
Ethanolic extract
200mg/kg |
3.93 ±1.13** |
45.71 ± 2.48** |
21.34 ±1.65** |
3.56 ± 2.96** |
|
5 |
Ethanolic extract
400mg/kg |
2.03 ± 0.45** |
33.21 ± 2.32** |
15.50 ±2.14** |
1.85 ± 1.08** |
Value represents mean ± SEM of six animals
in each group, Significant difference from the control, *P<0.05,
**P<0.01.
2.2.
Preparation of the extracts:
The collected roots were shade dried and powdered. The dried powder of the root of Cochlearia
armoracia was subjected for cold maceration with water and 70% ethanol for
5 days with occasional shaking and heating. At the end of fifth day both were
filtered through Buchner funnel. The filtrates were concentrated by vacuum
distillation. The concentrates were kept in petriplates and dried in freeze
drier until free from moisture. It was than weighed and kept in vacuum
desiccators to free from moisture and was used for further studies. The yield
for aqueous and ethanolic extracts was 20.8% and 18.6% w/w respectively.
Healthy adult wistar rats, weighing 180-250
g, were obtained from Animal house, JSS College of Pharmacy, Ooty, India and
maintained under standard laboratory conditions. The experimental protocol was
approved from Institutional Animal Ethical Committee (IAEC). The animal
experiments were carried out as per CPCSEA guidelines and after IAEC approval.
Renal damage was
induced in rats by 5/6 nephrectomy method according to the earlier reported
procedure (Podjarny et al., 2004; Braunlich et al., 1997). All
animals were allowed free acess to tap water and pellet diet and maintained at
room temparature in plastic cages.
2.5.
Experimental design:
The rats were
divided into five groups, consisting six animals in each group.
Group
1 Control group, treated with saline (2 ml/kg, p.o) daily for 15 days
Group 2 Treated with aqueous
extract 200 mg/kg b.w. daily for 15 days
Group 3 Treated with aqueous
extract 400 mg/kg b.w. daily for 15 days
Group 4 Treated with ethanol
extract 200 mg/kg b.w. daily for 15 days
Group 5 Treated with ethanol
extract 400 mg/kg b.w. daily for 15 days
At
the end of the treatment urine was collected by metabolic cages and blood
samples were collected through tail vein. The urine volume and the excretion of
metabolites, sodium and potassium ions were estimated according to the earlier
procedure (Afzal et al., 2004; Ganapathy et al., 2002).
Blood samples were collected in sterile centrifuge tubes allowed to clot. Serum
was separated and estimated for creatinine, urea, blood urea nitrogen and uric
acid (Englert and Harish, 1992).
2.6. Statistical
analysis:
The values are
represented as mean ± SEM. The results
were analyzed statistically by one-way analysis of variance (ANOVA), followed
by Dunnett’s post hoc test. “p” value of <0.01 was taken considered to be
significance.
The effect of the
root of Cochlearia armoracia on urinary volume and excretion of sodium
and potassium ions was given in Table 1.
Treatment with aqueous and ethanolic extracts of Cochlearia armoracia
showed significant increase in urinary volume and excretion of sodium and
potassium ions. Animal received only saline after nephrectomy (Control) had
shown much decrease in urinary volume and excretion of sodium and potassium
ions. The effect of the root of Cochlearia armoracia on Serum creatinine, urea, blood urea nitrogen and
uric acid was given in Table 2.
Treatment with aqueous and ethanolic extracts of Cochlearia armoracia
showed significant decrease in the levels of Serum Creatinine, Urea, Blood urea
nitrogen and Uric acid. Animal receives only saline after nephrectomy (Control)
had showed much increase in creatinine,
urea, blood urea nitrogen and uric acid concentration in serum. Aqueous
and ethanolic extracts reverse the effect of the renal failure caused by
nephrectomy. Aqueous and ethanolic extracts protected rats by lowering creatinine, urea, blood urea nitrogen and
uric acid concentration in serum after 15 days of the treatment. Aqueous
and ethanolic extract (400mg/kg) shows marked decrease in creatinine, urea, blood urea nitrogen and
uric acid levels as well as increase the urinary volume and excretion of
sodium and potassium ions.
4. CONCLUSION:
On the basis of
the result obtained, it can be concluded that the aqueous and ethanolic extract
of the roots of Cochlearia armoracia seems to possess renal protecting
activity in the rats. The results seem to support the traditional use of this
plant in renal protection.
The authors are grateful to
the management, JSS College of Pharmacy, for providing the necessary
infrastructure to carry out this research work in a successful manner.
1. Grieve M. A Modern Herbal. Dover Publication Inc, New York. 1995.
2. Bown D. Encyclopedia of herbs and their uses. DK Publishing Inc, New
York. 1996.
3. Hansen L. Content of glucosinolates in Horseradish(Armoracia Rusticana).
Tidsskr Planteavl. 1974; 73: 408-410.
4.
Chopra RN, Nayar SL,
Chopra IC. Glossary of Indian Medicinal Plants. Council of Scientific and
Industrial Research, New Delhi. 1992.
5. Goldberg A, Busse WR, Bluementhal M. A
therapeutic guide to herbals: The commission E Monographs. Boston, 1998.
6. Podjarny E, Hasdan G, Bernheim J, Rashid G,
Green J, Korzets Z et al. Effect of chronic tetrahydrobiopterin supplementation
on blood pressure and proteinuria in 5/6 nephrectomized rats. Nephrol Dial Transplant.
2004; 19: 2223-27.
7. Braunlich H, Marx F, Fleck C, Stein G. Kidney function in rats after
5/6 nephrectomy (5/6 NX): effort of treatment with vitamin E. Exp Toxicol
Pathol. 1997; 12: 135-139.
8. Afzal M, Iqbal A, Inamuddin M.
Diuretic and nephroprotective effect of Jawarish Zarooni Sada- a
polyherbal unani formulation. J
Ethnopharmacol. 2004; 91: 219-223.
9. Ganapathy S, Dash GK, Subburaju T, Suresh P. Diuretic, laxative and
toxicity studies of Cocculus hirsutus aerial parts. Fitoterapia 2002; 73:
28-31.
10. Englert J, Harmish Feger G. Diuretic action of aqueous Orthosiphon
extract in rats. Planta Medica. 1992; 58; 237-238.
Received on 04.02.2010
Accepted on 17.02.2010
© A&V Publication all right reserved
Research J. Pharmacology and
Pharmacodynamics. 2(4): July-August 2010, 300-302