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Ensete Superbum Ameliorates Renal Dysfunction in Experimental Diabetes Mellitus

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Ensete Superbum Ameliorates Renal Dysfunction in Experimental Diabetes Mellitus Iranian Journal of Basic Medical Sciences ijbms.mums.ac.ir Ensete superbum ameliorates renal dysfunction in experimental diabetes mellitus MS Sreekutty 1, S Mini 1* 1 Department of Biochemistry, University of Kerala, Kariavattom, Thiruvananthapuram‐695581, Kerala, India A R T I C L E I N F O A B S T R A C T Article type: Objective(s) Short communication : Hyperglycemia mediated oxidative stress plays a key role in the pathogenesis of diabetic complications like nephropathy. In the present study, we evaluated the effect of ethanolic extract of Article history: Ensete superbum seeds (ESSE) on renal dysfunction and oxidative stress in streptozotocin‐induced Received: Mar 14, 2015 diabetic rats. Accepted: Jul 23, 2015 Materials and Methods: Glucose, HbA1c, total protein, albumin, renal function markers (urea, uric acid and creatinine), and lipid peroxidation levels were evaluated. Renal enzymatic and non‐enzymatic Keywords: antioxidants were examined along with renal histopathological study. Antioxidant Results: ESSE (400 mg/kg BW t) administration reduced glucose and HbA1c, and improved serum total Diabetes mellitus protein and albumin in diabetic rats. ESSE in diabetic rats recorded decrement in renal function Diabetic nephropathy markers and renal lipid peroxidation products along with significant increment in enzymatic and non‐ Ensete superbum enzymatic antioxidants. Renal morphological abnormalities of diabetic rats were markedly Nephroprotection ameliorated by E.superbum. Streptozotocin Conclusion: These results suggest that the antioxidant effect of E. superbum could ameliorate oxidative stress and delay/prevent the progress of diabetic nephropathy in diabetes mellitus. ►Please cite this article as: Sreekutty MS, Mini S. Ensete superbum ameliorates renal dysfunction in experimental diabetes mellitus. Iran J Basic Med Sci 2016; 19:111‐118. Introduction complications of diabetes, nephropathy can be Diabetes Mellitus (DM) is a debilitating and often improved by antioxidants (4, 5). life‐threatening disorder with increasing incidence The major complication of diabetes is diabetic throughout the world (1). It is a chronic metabolic nephropathy, a leading cause of end‐stage renal failure disease characterized by hyperglycemia, resulting from in many developed countries, and a condition that insufficient or inefficient insulin secretion, with changes accounts for significant morbidity and mortality. In in carbohydrate, protein and lipid metabolism (2). diabetes, renal dysfunction develops through a number Diabetes and diabetes‐related complications represent of metabolic pathways, characterized by functional as one of the most central health problems worldwide, well as structural abnormalities of the kidneys. It is and according to recent estimations, it is likely to get characterized by a deterioration of the renal function worse to critical levels in the next decades, with the and changes in the glomerular structure, including great concern that this disease is rising rapidly in thickening of basement membrane, glomerular children and adolescents (3). hypertrophy and mesangial expansion (6‐8). Chronic hyperglycemia is a crucial factor in the Several mechanisms have been postulated for the development of diabetic complications such as kidney progression of diabetic nephropathy including diseases, heart diseases, Archive retinopathy and neuropathy. advanced glycation end products accumulation that of SID Direct or indirect consequence of hyperglycemia‐ stimulate mesangial cells to produce extracellular mediated overproduction of reactive oxygen species matrix (ECM), oxidative stress, acceleration of the (ROS) is the common pathophysiology shared by polyol pathway, and hemodynamic changes. Markers of microvascular complications of diabetes. Microvascular oxidative stress and reduced levels of antioxidants deterioration is avertable either by the inhibition have been found in tissues and/or blood, including of superoxide accumulation or by modulating the kidney, in both human and experimental animals in blood glucose levels and among the microvascular diabetes (9, 10). *Corresponding author: S Mini. Department of Biochemistry, University of Kerala, Kariavattom, Thiruvananthapuram‐695581, Kerala, India. Tel: +91‐471‐ 2308078; Fax: +91‐471‐2308078; email: [email protected] www.SID.ir Sreekutty and Mini Ensete superbum ameliorates renal dysfunction in diabetes Therefore, interventions favoring the ROS The weighed seed powder was extracted with scavenging and/or depuration (dietary and ethanol. The whole extract (ESSE) was collected, pharmacological antioxidants) prevent or attenuate filtered and further concentrated in vacuum under the oxidative stress, thereby ameliorating against the pressure using rotary flash evaporator. The dried subsequent renal damage (11). extract was suspended in distilled water and used for Current conventional therapies of diabetes using experimental study. blood glucose‐lowering medications have restrictions in averting the development of renal Chemicals diseases. Presently, research to develop drugs that All chemicals and biochemicals used in this study slow the progression of diabetic kidney damage with were of analytical grade and obtained from Sigma‐ fewer side effects is being conducted, however, Aldrich (St. Louis, MO, USA), Merck chemical showing no significant outcome (12). This has led to company (Darmstadt, Germany) and Sisco Research increasing consideration of complementary and Laboratories (Mumbai, India). alternative medicine from natural sources having potent antidiabetic as well as nephroprotective effect Experimental animals with fewer side effects. Normal healthy male Wistar albino rats (220–240 Indian rural and folklore ethnomedicinal g) were used for this study. The animals were housed practices include usage of numerous relatively in polypropylene cages in a room with temperature unidentified medicinal plants with scientifically non‐ maintained at 25 ±2 °C and a 12:12 hr light and dark characterized pharmacological activities. One such cycle. Animals were fed with laboratory chow less exploited folklore plant is Ensete superbum (Hindustan Lever Limited Lab diet) and water ad popular in Western Ghats of India, which is libitum. The protocol of this study was approved by consumed as an anti‐diabetic therapeutant by tribes Institutional Animal Ethics Committee. and by the local populace. E. superbum has been reported to have a broad range of therapeutic and Experimental induction of diabetes nutritional values. Diabetes was induced in overnight fasted rats by E. superbum (Roxb.) Cheesm., (Wild/Rock a single intraperitoneal injection of freshly prepared Banana) belongs to the family Musaceae, commonly Streptozotocin (STZ, 40 mg/kg body weight) in 0.1 M known as ‘cliff banana'. Indigenous communities citrate buffer of pH 4.5 (20). Rats were given 5% of consume its flowers, fruits and stem as a vegetable glucose in drinking water for the first 24 hr to (13‐15). In Ayurvedic system of medicine, the encounter any initial hypoglycemia. Animals were pseudostem and seeds of E. superbum were used for allowed free access to feed and water after the the treatment of various human ailments like injection. Hyperglycemia was allowed to develop diabetes, kidney stone (16), leucorrhea (17), measles over a minimum period of 48 hr. After 48 hr, animals (18), and stomach ache (19). However, there were no showing marked hyperglycemia (RBG>250 mg/dl) records of systematic pharmacological studies that were selected for the study. support this claim. Though there is no scientific evidence for the antidiabetic and nephroprotective Experimental design effects of E. superbum, tribal people have been using Rats were divided into four groups comprising six this plant in the management of DM. Preliminary in rats in each group. Group I was normal control rats; vitro studies revealed the antioxidant potential of E. group II consisted of normal rats treated with E. superbum seeds. The aim of the present study is to superbum seed extract at a dose of 400 mg/kg body ascertain the scientific basis for the use of E. weight; group III consisted of STZ‐induced diabetic superbum in the management of diabetes. Keeping in rats; group IV consisted of STZ‐induced diabetic rats view the protective effect of E. superbum in DM, the treated with ESSE at a dose of 400 mg/kg body present study was undertaken to explore the weight. On the fourth day after STZ injection, ESSE nephroprotective activity Archive of E. superbum seeds in treatment was started and th of SIDis was considered as the streptozotocin‐induced diabetic rats. first day of treatment. Intragastric administration of ESSE was continued for 60 days. At the end of the Materials and Methods experimental period, the rats were fasted overnight Plant material and extraction and sacrificed. Blood was collected in clean, dry test Seeds of E. superbum were collected from tubes and centrifuged at 3000 rpm for about 10 min Wayanad, Kerala, India and authentically identified to obtain blood serum. Serum samples were by Dr Valsaladevi (Curator, Department of Botany, separated immediately and used for the analysis of University of Kerala, Kerala, India). The seeds were serum biochemical parameters. Kidneys were dried and then powdered by a mechanical grinder. collected in ice‐cold containers for analysis of The hard outer coverings of seeds were removed. various biochemical parameters. 112 Iran J Basic Med Sci, Vol. 19, No. 1, Janwww.SID.ir 2016 Ensete superbum ameliorates
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