proposal for paper Essay

EVALUATION OF PROTECTIVE POTENTIAL OF ETHYL ACETATE

EXTRACT OF COCUS NUCIFERA IN GENTAMYCIN INDUCED

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

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

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

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

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

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

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

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

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]

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

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

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

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.

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 +

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.

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

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

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

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)

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

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

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

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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