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Nephroprotective and Nephrocurative Activity of Alangium Salvifolium Against Gentamicin Induced Nephrotoxicity in Albino Rats

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Nephroprotective and Nephrocurative Activity of Alangium Salvifolium Against Gentamicin Induced Nephrotoxicity in Albino Rats Karra Geetha et al. / Journal of Pharmacy Research 2014,8(4),576-583 Research Article Available online through ISSN: 0974-6943 http://jprsolutions.info Nephroprotective and nephrocurative activity of Alangium salvifolium against Gentamicin induced nephrotoxicity in albino rats Karra Geetha*1, Nadenla Ramarao 2 1CMR College of Pharmacy 1, Kandlakoya, Medchal, Hyderabad, Andhra Pradesh, India 2Chalapathi Institute of Pharmaceutical Sciences 2, Lam, Guntur, Andhra Pradesh, India Received on:09-03-2014; Revised on: 26-03-2014; Accepted on:11-04-2014 ABSTRACT Objective:To study the nephroprotective, nephrocurative effect of Alangium salvifolium ethanolic bark extract in gentamicin induced nephrotoxicity. Materials and methods: Nephrotoxicity was induced in wistar male rats by intraperitoneal administration of gentamicin at 40mg/kgb.wt /day for 21 days. Alangium salvifolium was selected to check the effect by using ethanolic bark extract with different doses (250,500,750 mg/kg body weight respectively), was given by oral route. Serum parameters (serum creatinine, serum urea,serum proteins, and blood urea nitrogen (BUN)), urine parameters (urine creatinine and urine volume) and other parameters like body weight, in vivo antioxi- dants catalase, reduced glutathione (GSH) and Lipid peroxidase level were determined on 22nd day in wistar male rats .Histopathological study of kidney was studied. Results: The three doses of the extracts produced significant nephroprotective, nephrocurative activities with increased doses. The increased actions of nephroprotective, nephrocurative activity in gentamicin induced nephrotoxicity models as evident by decrease in serum creatinine, serum urea,serum proteins, urine creatinine, BUN levels and lipid peroxidation (MDA).The increased glutathione (GSH), catalase (CAT) activities when compared to gentamicin control group which was further confirmed by histopathological study. Conclusion:The study revealed that ethanolic bark extract of Alangium salvifolium reversed the nephrotoxicity induced by gentamicin experimental animals. This indicates that Alangium salvifolium bark ethanolic extract can be used as an adjuvant with gentamicin to get the therapeutic benefit of these drugs without chances of its prominent nephrotoxicity side effect. KEYWORDS: Alangium salvifolium, nephroprotective, nephrocurative, antioxidants, gentamicin. 1. INTRODUCTION Drugs were shown to cause nephrotoxicity effects by one or more duced dose-dependent nephrotoxicity in 10-20% of therapeutic common pathogenic mechanisms. Drug-induced nephrotoxicity tends courses [3]. Gentamicin generates hydrogen peroxide in rat renal cor- to be more common among certain patients and in specific clinical tex mitochondria and it can also enhance the generation of reactive situations. Therefore, successful prevention requires knowledge of oxygen species (ROS) [4]. Abnormal production of ROS may damage pathogenic mechanisms of kidney injury, drug-related risk factors, some macromolecules to induce cellular injury, direct tubular necro- patient-related risk factors, and preventive measures, coupled with sis, without morphological changes in glomerular structures [5, 6]. Pos- vigilance and early intervention [1, 2]. sible gentamicin mechanisms to induce nephrotoxicity are peroxidation of membrane lipids, protein denaturation and DNA damage [7-9].Gen- A number of antibiotics including the tetracyclines, cephalosporins, tamicin also acts as an iron chelator and the iron-gentamicin complex penicillins, as well as aminoglycosides and sulfonamides, are poten- is a potent catalyst of free radical generation. tial nephrotoxins. Gentamicin, a typical aminoglycoside antibiotic is widely used in clinical practices for the treatment of life threatening Alangium salvifolium(L.f) Wang belongs to family Alangiaceae. Lo- gram-negative infections. This antibiotic generally causes drug-in- cally it called as Ankolam. Alangiaceae is a monogeneric family of trees and shrubs found in tropical and subtropical region.The plant is distributed in dry regions, plains and lower hills in India, Africa, Srilanka, and China. *Corresponding author. K.Geetha Alangium salvifolium wang was medicinally used in India, China and CMR College of Pharmacy, Kandlakoya (v), Medchal(M), Philippines. The parts of the plant are used to treat different diseases. Hyderabad, Andhra Pradesh, India. Root bark was used as an antidote for rabies. Fruits are sweet and it’s Journal of Pharmacy Research Vol.8 Issue 4.April 2014 576-583 Karra Geetha et al. / Journal of Pharmacy Research 2014,8(4),576-583 used to treat burning sensation, constipation and haemorrhage. Stem Group-I (Normal control): received saline (1ml/kg b.wt, p.o). bark exerts a biphasic action on the blood pressure in cats at lower Group II (Toxic control): received gentamicin (40mg/kg b.wt for doses and marked hypotension in higher doses. 21days i.p). Group III (Plant control): received plant extract (200mg/kg b.wt for The plant has been reported for its antitubercular, antispasmodic, 21 days p.o). antiarthritis, antibacterial, anti fungal and anticholinesterase activity. Group IV (prophylactic): received gentamicin (40mg/kg b.wt, i.p) + This plant was used as antirheumatic agent by the local people of 250mg/kg b.wt of plant extract (p.o). Vellore and Tirupattur districts in Tamilnadu. Root bark was used as Group V (prophylactic): received gentamicin (40mg/kg b.wt, i.p) + an anthelmintic, antiemetic, febrifuge, hypoglycemic, purgative, 500mg/kgb.wt of plant extract (p.o). antileprotic agent and other skin diseases. The seed oil was used Group VI (prophylactic): received gentamicin (40mg/kg b.wt, i.p) + medicinally for externally and internally with palm jaggery for syphi- 750mg/kg b.wt of plant extract (p.o). litic ulcers and scabies, gonorrhea and internally for leprosy. Stem bark used to control vomiting and diarrhea. Previously ethanol ex- The IV,V, VI Prophylactic group animals were treated with gentami- cin (40mg/kg b.wt, i.p),1 hr prior to administration of Alangium tract has been reported to possess anti-microbial,analgesic and anti- salvifolium bark ethanolic extract of respective doses for 13 days and inflammatory activities [10-17]. only plant extract of respective doses will be given from 14th-21st day. 2. MATERIAL AND METHODS Group VII (Curative Group): received gentamicin (40mg/kg b.wt, i.p) for 13 days and best dose of the plant extract from the prophylactic 2.1. Collection of plant materials: treatment was selected and will be given from 14th day to 21stday. Alangium salvifolium bark was collected from Hyderabad city and authenticated by P.V.Prasanna. (Scientist / officer In-charge) 2.5. Sample Collection: Botanical Survey of India bearing no. BSI/DRC/12-13/Tech./736. Rats were placed in metabolic cages over a period of 24hr and urine volume collected was measured and it was used for estimation of 2.2. Preparation of Alangium salvifolium extract: creatinine. The bark was allowed to dry under shade. The dried bark was pow- ered in a mill and extracted successively by using various solvents Animals were sacrificed on 22nd day of the study after collection of like n-hexane, ethyl acetate, ethanol, water. blood by retro orbital puncture and were centrifuged to separate the serum. It was used for estimation of creatinine, blood urea nitrogen Required quantity of ethanolic bark extract of Alangium salvifolium and total proteins. 250, 500,750mg/kg body weight of the rat was weighed and adminis- tered. 2.6. Parameters assessed: 2.3. Animals Body weight: Healthy wistar adult male albino rats weighing about 150-200gm were The weight of the animals (in grams) was noted on the 1st day and last housed in polypropylene cages and maintained at 24± 2 ºc under 12 day of the study and the difference in body weights was noted. hr light/dark and 60±5% humidity. They were fed with standard rat pellet diet and water ad libitum. Animals were acclimatized to our lab Serum creatinine: environment for about a week under laboratory conditions. All ex- Serum creatinine level was estimated by alkaline picrate method us- periments were performed according to the ethical standards of ani- ing creatinine kit and read absorbance at 520nm. mal handling and approved by Institutional Animal ethics committee (CPCSEA/1657/IAEAC/ CMRCP/PhD-l3/11-A). Blood urea Nitrogen (BUN): BUN level in serum was estimated by kit of Auto span Pvt Ltd. and 2.4. Experimental Protocol read the absorbance at 570nm. Fifty six male wistar rats were used for the study and animals were Total Proteins: divided into seven groups containing eight animals in each. Total proteins level in serum was estimated by kit of Auto span Pvt Ltd. and was read at 578nm. Journal of Pharmacy Research Vol.8 Issue 4.April 2014 576-583 Karra Geetha et al. / Journal of Pharmacy Research 2014,8(4),576-583 Urine volume: Volume of urine was measured in all groups during last 24 hrs of compared to gentamicin control group. The data was analyzed statis- experiment in all groups. tically by one way ANOVA followed by Dunnett test using graph pad prism 5.0 Software. Values are expressed as mean ± SEM (n =8). Where Urine creatinine: Urine creatinine was estimated by alkaline picrate *** (p<0.001)vs gentamicin control, ### (p<0.001) vs normal control. method using creatinine kit and read the absorbance at 520nm. 3. RESULTS GSH: GSH level in the kidney homogenate was estimated by the method of 3.1. Effect of Alangium salvifolium bark ethanolic extract on body Ellman.et al.1959. [18] weight in gentamicin induced
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