EVALUATION OF PROTECTIVE POTENTIAL OF ETHYL ACETATE
EXTRACT OF COCUS NUCIFERA IN GENTAMYCIN INDUCED
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NEPHROTOXICITY IN ALBINO MICE.
Abstract: Renal disorders have always remained a major area of concern for physicians
since a long time and most are of these are drug induced . Antibiotic are used to treat
infections caused by organisms that are sensitive to them , e.g gentamycin which is an
aminoglycosides which are ototoxic and nephrotoxic . This study looks at the
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Nephroprotective activity of ethyl acetate ext ract of husk fibre of Cocus nucifera for its
protective effects on gentamicin -induced nephrotoxicity in albino mice.
For studying
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acute toxicity study, the study groups contained eight rats in each group and oral dosage
of 100, 50, 25mg ethyl acetate husk fibre of Cocus nucifera extract/kg body weights was
administered to albino mice. Nephrotoxicity was induced in albino mice by daily
intraperetoneal administration of gentamicin 45 mg/kg/day for 10 days. Effect of
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concurrent administration of ethyl acetate extract of Cocus nucifera at a dose of 100, 50
and 25 mg/kg/day given by oral route for 17days.
The biochemical parameter such as
serum creatinine, uric acid, blo od urea nitrogen and serum electrolytes as indicators of
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kidney damage was determined by using one way ANOVA the results are significant at
P> 00 5. the result shows that at higher concentration of the extract, kidney damage was
not seen while lower concentration it was seen.
INTRODUCTION:
Nephrotoxicity is one of the most common kidney problems and occurs when
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body is exposed to a drug or toxin. When kidney damage occurs, the body unable to rid
of excess urine and wastes from the body and blood electrolytes (such as potassium and
magnesium) will all become elevated and Sodium will reduce . A number o f therapeutic
agents can adversely affect the kidney resulting in acute renal failure, chronic intestinal
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nephritis and nephritic syndrome because increasing number of potent therapeutic drugs
like aminoglycoside antibiotics, chemotherapeutic agents and NS AIDs have been added
to the therapeutic arsenal in recent years. Exposure to chemical reagents like ethylene
glycol, carbon tetra chloride, sodium oxalate and heavy metals like lead, mercury, arsenic
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and cadmium also induces nephrotoxicity. [1]
Many plants have been used for the treatment of kidney failure in traditional
system of medicine throughout the world. Indeed along with the dietary measures, plant
preparation formed the basis of the treatment of the disease until the introduction of
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allopathic medi cine. Traditional knowledge will serve as a powerful search engine and
most importantly, will greatly facilitate intentional, focused and safe natural products
research to rediscover the drug discovery process. Therefore, search of nephroprotective
herbs f rom medicinal plants has become importan t and need of the day. [2]
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Nephroprotective agents are the substances which possess protective activity
against nephrotoxicity. Medicinal plants have curative properties due to the presence of
various complex chemica l substances. Ancient literature has prescribed various herbs for
the cure of kidney disease. [3]
Coconut tree has been eulogised as Kalpavriksha (the all giving tree) in Indian classics
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for is all round usef ulness , [4] , Husk fibre of a coconut tress ha s been reported to have
antibacterial, antifungal, antiviral, antiparasitic, antidermatophytic, antioxidant,
hypoglycemic, hepatoprotective, immunostimulant, antiblenorrhagic, antibronchitis,
febrifugal, and antigingivitic properties, [4], and antimalaria l activity, [5] . Very few
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studies were reported in literature regarding histopathology of gentamicin i nduced renal
failure in Albino mice . So, the presen t study is taken up to see the effect of ethyl acetate
extract of cocus nicifera, it nephroprectective properties by investigating the biochemical
and histopathologica l changes o f mice .
Materials a nd Methods
Wistar albino mice weighing 20 -26gms, are utilized for the present study. Experiments
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were performed with the permission of the institutional ethics committee. In th e present
study, albino mice were used and are grouped as follows:
Group A: (Control Positive): Administered appropriate volume of 5% DMSO solution.
Group B: (Control Negative): Administered appropriate volume of 5% DMSO solution +
200?l Gentamycin.
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Group C: Administered 100mg/Kg body weight of Cocus nucifera extract fraction +
200?l Gentamycin.
Group D: Administered 50mg/Kg body weight of Cocus nucifera extract fraction + 200?l
Gentamycin.
Group E: Administered 25 mg/Kg body weight of Co cus nucifera extract fraction +
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200?l Gentamycin.
METHOD
All rats we re kept under observation for 2 week prior to the experiments to permit the
animals to adjust to the environment. All animals were fed standard rat chow and were
provided tap water to drin k ad libitum . They were housed in a facility with 12 12 h light
dark cycle that is maintained at 25°C. All animals were weighed before the injections.
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The anima ls were anaesthetized with diethyl ether inhalation. Blood samples were
collected with cardiac p uncture for biochemical investigations like blood urea, uric acid,
creatinine, serum Na, K, Cl and HCO 3 were determine.
Administration of the sample: daily intraperitonial injection of gentamycin was given
to each group for 10 days and the daily oral admi nistration of ethyl acetate extract of
cocus nucifera are given to each group for 17 days .
Collection of Blood Sample: At the end of the 17 -day experimental period, the Mice
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were sacrificed by slight diethyl ether anaesthesia, the neck area was quickly cle ared of
fur and the jugular veins exposed, from which blood was collected into EDTA bottle to
prevent clotting. The EDTA blood sample was centrifuged at 3000 rpm for 10 minutes
and the serum pipetted out. This was stored frozen at -20°C until needed for an alysis.
Determination of Kidney function test (serum urea, creatinine and uric acid): -The
serum parameters were analyzed spectrophoto -metrically by using Spectrumlab 752S
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UV VIS. Estimation of blood urea, uric acid and creatinine Assay kits for serum ana lysis
were obtained from Randox Laboratories Ltd, UK, according to the methods of Talke and
Schubert et al (1965), Tiffany et al (1972) respectively.
Determination of serum electrolytes: -Sodium (Na) and Potassium (K) analysis were
carried out using Randox Laboratory kit according to the method of (Terri A.E et al
1958) . Serum calcium was determined colorimetrically using commercial kits (Erba,
Germany) according to the meth od of Moorehead W R et al 1974[8 ].
Determination of serum Enzymes (ALP, AST, and AL T): – The serum parameters
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were analyzed spectrophoto -metrically by using Spectrumlab 752S UV VIS and assay
kits for serum enzymes analysis were obtained from Randox Laboratories Ltd, UK, ALP
was carried out by the method of Wright et al. (1972), The metho d described by Reitman
and Frankel (1957) was used to determine the activity of AST and ALT.
.
RESULTS
Table 1: Effect of polyphenols extract of Cocos nucifera husk fibre (WAT) on
selected kidney parameters of Gentamycin induced renal impairment in Mice
Group CREAT
(mg/dl)
BUN
(mg/dl)
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URIC ACID
(mg/dl)
Control (5% DMSO) 0.25±0.02 a 34.65±1.50 a 0.46±0.04 a
Gentamycin 0.34±0.02 b 48.95±2.72 b 0.99±0.07 c
Gentamycin + 25mg/kg body weight of extract 0.25±0.02 a 36.25±3.48 a 0.56±0.07 b
Gentamycin + 50mg/kg body weight of extract 0.29±0.03 a 37.22±1.28 a 0.69±0.08 b
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Gentamycin + 100mg/kg body weight of extract 0.31±0.02 b 41.28±1.21 a 1.12±0.07 c
Data are mean ±SEM of five determinations. Values with the same superscript are not
significantly different at p < 0.05
Table 2: Effect of polyphenols extract of Cocos nucifera husk fibre (WAT) on some
selected electrolytes of Gentamycin induced renal impairment in Mice
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Group Na +(Meq/L) K+(Meq/L) Cl – HCO 3
Control (5% DMSO) 156.08±4.42 a 4.50±0.33 a 91.25±2.5 b 26.15±1.62 a
Gen tamycin 147.85±2.64 b 7.50±0.32 b 82.00±6.58 b 23.02±2.33 a
Gentamycin + 25mg/kg
body weight of extract
151.05±0.78 a 5.37±0.21 a 78.00±5.59 a 24.45±1.67 a
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Gentamycin + 50mg/kg
body weight of extract
147.95±4.09 a 5.40±0.23 a 90.00±3.92 b 22.00±1.22 a
Gentamycin + 100mg/kg
body weight of extract
153.25±5.47 a 5.68±0.32 a 95.00±3.74 b 20.95±1.34 a
Data are mean ±SEM of five determinations . Values with the same superscript are not
significantly different at p < 0.05
Table 3: Effect of polyphenols extract of Cocos nucife ra husk fibre (WAT) on some
selected enzymes of Gentamycin induced renal impairment in Mice
Group ALT AST ALP
Control (5% DMSO) 20.00±2.94 a 39.00±4.50 a 12.58 ±1.8 3a
Gentamycin 16.00±2.94 a 32.00±3.92 a 8.83 ±1.25 a
Gentamycin + 25mg/kg bo dy
weight of extract
22.00±3.36 a 38.00±3.37 a 45.39 ±1.75 c
Gentamycin + 50mg/kg body
weight of extract
22.00±2.58 a 32.00±2.58 a 39.58 ±1.72 b
Gentamycin + 100mg/kg body
weight of extract
18.00±2.58 a 84.00±3.65 b 13.50 ±1.83 a
Data are mean ±SEM of five determinations . Values with the same superscript are not
significantly different at p < 0.05.
DISCUSSION
The incidence of renal dysfunction following amino -glycoside administration was
detected by many w orkers (Garetz and Schacht1996[9]; Baliga et a l., 1997[10] and Abdel
Naim etal.,1999[11 ]). The administration of gentamycin into mice induced impairment of
renal function through liberation of oxygen free radical (Heibashy & Abdel Moneim,
1999 [12] and Heibashy et al.,2009[13]). Renal failure is chara cterized by disorders in
some biochemical parameters and Kidney function indices . It can be seen that
gentamycin produce Nephrotoxicity in the mice by seen the increase in Creatinine,
Potassium and lowering the Sodium .
These results confirmed that gentamic in produced nephrotoxicity as previously reported
by Ali et al., 2003[14], Goto, 2004 [15] and Heibashy et al .,2009[13]. Serum e lectrolytes
were disturbed in gentamycin treated mice as compared with control animals and the
ethyl acetate of Cocus nucifera extract was seen to be able to attenuate this trend by
trying to correct the damages done. Those group that receive 100 and 50 mg/kg b.wt of
the extract are found to be doing well and the effectiveness of the extract was seen while
those that receive the 25 mg/kg b.wt, the renal function was seen to be serious in them.
Lower value of serum sodium indicated inability of kidney to conserve sodium and
chloride. Haemodilution too may be involved in the fall of sodium value via excess of
water intake and or incre ased production of endogenous water. Increase of Potassium
may be due to reduced excretion of K aggravated by leakage of intracellular potassium
into blood stream as a result of gentamicin induced lesions in renal tubular epithelium.
The present results ar e in harmony with the data obtained by Heibashy & Abdel Moneim
(1999)[12] and Heibashy et al. (2009)[13] .
V. Conclusion
Daily administration of ethyl acetate extract of cocus nucifera for 17days was seen to be
able to attenuate the renal dysfunction caus e by daily intraperitonial injection of
gentamicin 45mg/ kg b.w for 10 days is evident on renal function tests. Thus, it could be
suggested that gentamicin must be given in the lowest effective therapeutic doses in
patients with normal kidney function. Also , gentamicin therapy should be preceded by
antioxidant administration and also it could be suggested that the husk fibre of ethyl
acetate extract of cocus nucifera can be modified into drug so that it can be given before
administration of gentamycin becaus e it is seen from this presence stud ies that it has
nephroprotective properties
Acknowledgements
I really appreciate Dr . Adebayo , who took his time to go through this piece of work, also
to the Professor s and all the staff of the Department of Biochemist ry, University of Ilorin,
Kwara State, Nigeria .
Note: This st udy is a partial work of my PGD research , titled evaluation of protective
potential of ethyl acetate extract of Cocus nucifera in gentamycin induced nephrotoxicity
in albino mice .
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