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Protective Role of Excoecaria Agallocha L. Against Streptozotocin

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Protective Role of Excoecaria Agallocha L. Against Streptozotocin The Pharma Innovation Journal 2018; 7(9): 17-26 ISSN (E): 2277- 7695 ISSN (P): 2349-8242 NAAS Rating: 5.03 Protective role of Excoecaria agallocha L. against TPI 2018; 7(9): 17-26 © 2018 TPI streptozotocin induced diabetic complications www.thepharmajournal.com Received: 10-07-2018 Accepted: 12-08-2018 G Kiran, Ganesh N Sharma, Birendra Shrivastava and AMS Sudhakar Babu G Kiran School of Pharmaceutical Abstract Sciences, Jaipur National Oxidative stress has a pivotal role in the pathogenesis and development of various diabetic-related University, Jagatpura, Jaipur, complications, the most common and debilitating effects of diabetes mellitus are on kidney, liver and India heart tissue. There is accumulating evidence that Excoecaria agallocha L extract, a rich source of phenolic and flavonoid components, has hypoglycemic and antioxidative properties. This study aimed to Ganesh N Sharma determine the protective effects of Excoecaria agallocha L extract against streptozotocin-induced School of Pharmaceutical diabetic complications on kidney, liver and heart in rat. The diabetic rat model was generated by Sciences, Jaipur National intraperitoneal injection of streptozotocin (STZ) at a dose of 60 mg/kg, i.p. dissolved in 0.01M citrate University, Jagatpura, Jaipur, buffer (pH 4.5) after 16 h fasting. A subset of the STZ-induced diabetic rats intragastrically administered India with Excoecaria agallocha L. extract (250 and 500 mg/kg/day) before or after the onset of diabetes for Birendra Shrivastava 16 weeks, whereas other diabetic rats received only isotonic saline as the same volume of Excoecaria School of Pharmaceutical agallocha L extract. To evaluate the effects of Excoecaria agallocha L extract on the diabetic rats Sciences, Jaipur National various parameters, including oxidative parameters, liver enzymes, kidney-specific parameters, and University, Jagatpura, Jaipur, histopathology were assessed along with other biochemical assessments. Greater blood sugar, SGOT, India SGPT, creatine kinase, urea whereas lower uric acid, creatinine and total protein were detected in the STZ-diabetic rats, which were significantly attenuated after Excoecaria agallocha L extract AMS Sudhakar Babu administration, dose-dependently. Even various oxidative stress parameters were also drastically changed A.M Reddy Memorial College of in a STZ-diabetic rats which were improved by Excoecaria agallocha L extract. Histopathological study Pharmacy, Narasaropet, Andhra of liver, kidney and heart tissue were supported the potential protective effect of Excoecaria agallocha L Pradesh, India extract. The activity elicited by the extract might be due to its ability to activate antioxidant enzymes. The findings suggest the potential use of the extract of Excoecaria agallocha L. as a novel therapeutically useful nephron protective, hepatoprotective and cardio protective agent. Keywords: Excoecaria agallocha L, STZ, antioxidant, hepatoprotective, nephron protective, cardio protective 1. Introduction Diabetes Mellitus is a constitutional disease. It is also known as “Disease of Civilization” (Urbanization) seen more in cities than villages. But however, it is now making inroads into Indian villages. Diabetes is an important human ailment afflicting many from various walks of life in different countries. It is an outcome of sedentary lifestyle & incorrect food habits. No. of people afflicted by Diabetes Mellitus is increasing each day. 9-12% population of world has either established Diabetes Mellitus (or) tendency of contracting it in near future [1, 2].18 Millions of Indians are suffering from Diabetes Mellitus. Though it is rampant today, it is not a new disease. It is well-known from historic times. Well known ayurvedic physicians Maharshi Charaka (600BC) & Sushrutha (400BC) correctly described almost all the symptoms of this disease and called it as “Madhu Meha” (a shower of honey) and explained in Ayurvedic literature called ‘Sushrutha Samhitha’ [3-6]. Nature has provided us plenty of medicinal plants and in fact, all of them have not been explored yet. The medicinal value of plant lies in some chemical substances that produce a definite physiological action on the human body. The most important of these bioactive components of the plants are alkaloids, flavonoids tannins, and phenolic compounds. These are reports that plant possessing phenolic compounds, flavonoids and tannins are useful as antioxidants [7, 8]. Keeping these points in view, Excoecaria agallocha L. is a mangrove species that belongs to Euphorbiaceae family and widely occurs on the south-east coast of India. Correspondence Traditionally, Excoecaria agallocha L was used to treat sores and stings from marine G Kiran creatures, for treating ulcers, as a purgative and an emetic, and the smoke from the bark to treat School of Pharmaceutical leprosy Scientifically E. agallocha was reported to have antifilarial, antioxidant, Sciences, Jaipur National cytoprotective, antifouling, antinociceptive and piscicidal activities [9-11]. University, Jagatpura, Jaipur, India ~ 17 ~ The Pharma Innovation Journal This is due to the presence of diterpenoids, triterpeneoids, Solution B flavonoid and phorbole esters [12-14]. The leaves of the plant 200 mg of powder is dissolved in water, filtered and distilled have rich in polyphenols and flavonoids due to the presence water is added, heated for few minutes, 0.1 ml of Fehling’s A of these constituents the plant may be has antioxidant and B solution is added. Heated for 2 minutes, red precipitate property. Diabetic Mellitus is usually conveyed by excessive is formed. production of free radicals, hyperglycemia-induced Then compared the solution B with A. mitochondrial reactive oxygen species production could be a key episode in the progress of diabetic complications [15-17]. Borntrager’s test: A few ml of dilute sulphuric acid is added The antioxidant property of the plant has deteriorates the to the extract solution, boiled, filtered and treated the filtrate progression of diabetic-induced complications. with chloroform and shaken well. Then separated the chloroform layer and tested with a few ml of ammonia Material and Methods solution. The ammoniacal layer formed pink or red color. Identification and Authentication of Plant Materials The plant Excoecaria agallocha L. identified and Modified Borntrager’s test: Few ml of 5% Fecl3 solution authenticated by Dr. Madhava Chetty, department of botany, and dilute hydrochloric acid is added to the extract. Then S.V. University, Tirupati and preserved in herbarium for boiled for 5 min, cooled and shaken well with organic solvent. further identification. Then added equal quantity of dilute ammonia. The ammoniacal layer formed pinkish red color. Collection of Plant Materials The leaves of Excoecaria agallocha L. collected in the month Legal’s test: Extract is dissolved with pyridine and made of April – May at coastal region of Andhra Pradesh. The alkaline with sodium nitroprusside solution. The solution collected materials are washed and dried in shade. The dried becomes pink or red. materials are powdered by using a mixer. The powdered drug is subjected to solvent extraction by soxhlet apparatus. Ballet's test: 1 ml of sodium picrate solution is added to the extract. Color changes from yellow to orange. Extraction Procedure Keller-killani test: Powdered drug extract is added to the Fine powder of Excoecaria agallocha L. is subjected to sox 10ml of 70% ethanol for 2 min, then filtered and added 10 ml elation by using 70% ethanol as solvent for a period of 72 of water, 0.5 ml of strong solution of lead acetate and filtered. hours at the temperature of 60 °C obtain dark colored extract The filtrates are shaken with 5 ml of chloroform and separate is cooled, filtered and concentrated by evaporation and freeze- the chloroform layer and evaporated. To this 3 ml of glacial dried. The extract thus obtained is used for further acetic acid is added and cooled. Then added 2 drops of 5% experimental studies. Dried extract is dissolved in distilled ferric chloride solution. Transferred the contents into a test water for animal administration [18, 19]. tube containing 2 ml of concentrated sulphuric acid. Reddish brown layer was formed at the junction of the two liquids and Preliminary Phytochemical Studies [20] [21] the upper layer slowly becomes bluish green on standing. Qualitative phytochemical analysis The extract obtained by solvent extraction method is C. Test for Flavonoids subjected to qualitative phytochemical analysis in order to Shinoda test: To the ethanolic extract of powder, magnesium identify the nature of constituents present in the prepared turnings or foil and conc. Hcl are added. Intense cherry red extract solution. color or orange-red color is formed. The detailed study about the phytochemical test procedure as follows General tests: To the small quantity of residue, lead acetate solution is added. Yellow color precipitate is formed. A. Test for carbohydrates Addition of increasing amount of sodium hydroxide to the Molisch’s test: 2-3 ml of extract is added with 1ml of freshly residue shows yellow coloration, which decolorizes after prepared alpha-naphthol solution in alcohol, shaken and addition of acid. added conc.H2SO4 from sides of the test tube under tap water. Violet ring is formed at the junction of two liquids. D. Test for Saponins Benedict’s test: To the extract solution, Benedict’s reagent is Foam test: Small quantity of ethanolic extract and 20 ml of added, heated on water bath and red color is formed.
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