EFFECT OF STANDARD GLYCLAZIDE DRUG AND INDIGENOUS COW URINE POWDER OVER HEPATIC GLYCOGENOLYSIS AND GLUCONEOGENESIS IN DIABETIC RATS

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EFFECT OF STANDARD GLYCLAZIDE DRUG AND INDIGENOUS COW URINE POWDER OVER HEPATIC GLYCOGENOLYSIS AND GLUCONEOGENESIS IN DIABETIC RATS
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  EFFECT OF STANDARD GLYCLAZIDE DRUG AND INDIGENOUS COW URINE POWDER OVER HEPATIC GLYCOGENOLYSIS AND GLUCONEOGENESIS IN DIABETIC RATS Yashwant Atbhaiya, Dr. P.L. Choudhary. Kamdhenu and Panchgavya Research centre, Durg. Email:- Yashwant.Atbhaiya@gmail.com  Abstract The experiment consists of six treatments which were replicated six times under complete randomize design. In this experiment seven treatments i. e; normal control (without diabetes), diabetic control (diabetes induced by streptozotocin 50 mg/kg body weight/ rats), standard drug group (Glyclazide 1.44 mg/ 200g body weight rats), cow urine powder (100 mg), cow urine powder (200 mg) and cow urine powder (400 mg). Proximate composition of cow urine powder maximum recorded creatinine (154.12 mg), potassium (164.27 mg), calcium (113.39 mg), sulphate (83.48 mg), total nitrogen (1.79 g), urea (21.94 g), total solids (93.50 %), moisture (6.50 %) and pH (8.1). In this experiment treatment combination of cow urine powder 400 mg group of rats which was streptozotocin treated and observed the glucose level and lipid profile level upto 15 th  days. Keywords: Antidiabetic, skim milk powder, streptozotocin, glyclazide, lipid profile. Introduction In today’s world health scenario there are many factors driving the growth of diabetes,  but most experts agree that changes in lifestyle and diet are the major attributes. As developing countries rapidly industrialize, people tend to do work involving less physical activity. At the same time, the availability of food that is cheap but high in calories becomes more common. Stress and other psychological phenomena also play a key role in widespread illnesses. The trend emerging is that the age ranges have been dropping, so that people are getting sick in the prime of their life. Diabetes mellitus is certain to be one of the most challenging health problems in the 21 st  century. It is most common endocrine disorder caused by absolute lack of insulin or a relative lack of insulin. All over the world it is the important multifactor commonest  metabolic disorder in men and women. Recently this could be expected to be 380 million. Its incidence has been estimated to be around 15% of Indian population. World Health Organization (WHO) has projected India as the leading country in the world, as per diabetic concerned. In the year 2025 the diabetic population in India will reach up to 70 million. In the  present day life style people are more prone to disease in general and more to metabolic disorders. It is epidemic in many developing and newly industrialized nations. It is major global health problem with diverse causative factors often associate with multiple devastating innervating complications, increasing disability and reduced life expectancy. The information available in the classics of Ayurveda shows that diabetes mellitus as a disease was very well known to the propounders of Ayurveda (Pandey,   2010). Material and methods The first early morning urine samples were collected from the cow shed. The cow urine samples were dried at different temperature such as 45 0 C, 55 0 C and 63 0 C in the laboratory vacuum oven. Drying of cow urine samples at temperature 63 0 C was found to be more effective than that of other temperature. All chemicals and reagents used were of analytical grade and obtained from Himedia Laboratories Pvt. Limited, India. The kits for the estimation of blood glucose and serum lipid profile were obtained from Accue-check meter, Roche Diagnostics, USA and Lab-care Diagnostics Pvt. Ltd, India. The standard drug Glyclazide was purchased from Hyderabad, India. Other chemicals such as sodium citrate, HCl, NaOH, and other raw materials such as insulin syringe, needle, cotton, capillary tube and cages were purchased from local market. Selection and registration Albino Wistar rats of both sex (male and female) were procured from the laboratory of Columbia Institute of Pharmacy Tekari, Raipur 493111 (C.G.), India, and were maintained under a 12 h light/dark cycle and allowed for feed and water without limit. Approved experimental protocol in accordance with the guidelines provided by the committee for the control and supervision of experiments on animals (CPCSEA). The experimental rats were registered by CPCSEA and registration number was 1321/ac/10/CPCSEA. Induction of diabetes in different groups of rats using streptozotocin solution  Diabetes was induced in rats by the intra peritoneal injection of streptozotocin (Himedia) at a dose of 50 mg/kg body weight dissolved in citrate buffer (0.1 M, pH 4.5) in the volume of 1 ml/kg. The diabetic rats were given 5% w/v glucose solution orally in order to prevent hypoglycemia during the first day after the streptozotocin administration. Three days after the injection (six group), the blood glucose levels were measured and the animals with blood glucose levels above 300 mg/dl were consider being diabetic and were used in the subsequent experiments. In all the experiments, rats were fasted for 16 hr prior to streptozotocin injection. Experimental rats were divided in 6 groups of 6 rats per group. The details of treatments were given below: Group 1-  Normal control group (without diabetes). Group 2- Diabetic control group (streptozotocin-treated).  Group 3- Standard drug group- Diabetic animals received daily a single oral dose of the reference drug Glyclazide 1.44 mg/200 g body weight from day 1 to 15. Group 4 (a) - Diabetic animals received daily a single oral dose of cow urine powder (100 mg). Group 4 (b)  –    Diabetic animal received daily a single oral dose of cow urine powder (200 mg). Group 4 (c)  –    Diabetic animals received daily a single oral dose of cow urine powder (400 mg). Statistical analysis was done by using complete randomized block design (CRD)-single factor analysis of variance of the effect within and between the treatments groups at 1% level (Snedecor and Cochran, 1967) using SPSS software (Version SPSS, 1999, SPSS I User’s Guide; release 10.0.1ed. SPSS Inc. USA). Result and discussion Effect of cow urine powder on blood glucose, cholesterol, HDL, LDL, and triglyceride level in streptozotocin induced rats after 15 days of experiment and observed the overall effect of the treatment. The overall effect of glucose and lipid profile levels in diabetic rats treated with various control measures such as standard drug glyclazide and various levels of cow urine powder (100, 200, 400 mg) during the 15 days of experiment.  Results obtained during the treatment period for blood glucose level in various groups such as normal control, diabetic control, standard drug glyclazide, cow urine powder 100 mg, cow urine powder 200 mg and cow urine powder 400 mg were 91.66, 447.66, 125.25, 174.75, 163.25 and 133.50 mg/dl respectively. The lipid profile levels were determined during the experiment for all groups. In normal control group, diabetic group, standard drug group and cow urine powder group (100, 200 and 400 mg) contains cholesterol level was 81.65, 126.37, 67.72, 95.55, 93.70 and 90.82 mg/dl respectively. The value obtained for LDL level for various groups such as normal control group, diabetic control group, standard drug group, cow urine powder group 100 mg, cow urine powder group 200 mg and cow urine  powder group 400 mg were 28.11, 86.87, 16.17, 51.15, 47.22 and 40.85 mg/dl respectively. Similarly for HDL level for various groups such as normal control group, diabetic control group, standard drug group, cow urine powder group 100 mg, cow urine powder group 200 mg and cow urine powder group 400 mg were 53.53, 39.50, 51.55, 44.40, 46.47 and 49.97 mg/dl respectively. The sample also analyzed for triglyceride content such as normal control group, diabetic control group, standard drug group, cow urine powder group 100 mg, cow urine powder group 200 mg and cow urine powder group 400 mg were 77.21, 188.67, 95.35, 134.55, 132.70 and 126.47 mg/dl respectively. The data indicated that glucose level for normal control group was 91.6 mg/dl and streptozotocin was introduced in normal group and rats become diabetic and blood glucose level was 447.6 mg/dl. After induction of diabetes in the rats and treated with standard drug glyclazide and the blood glucose level was reduce from 599.1 to 125.2 mg/dl. Within 15 days of experiment. Similarly the other control measure such as various level of cow urine powder 100, 200 and 400 mg reduced significantly (p<0.01) from 632.0 to 174.7, from 547.6 to 164.6 and from 630.5 to 133.6 mg/dl respectively. The cholesterol level for normal control group is 81.65 mg/dl whereas in diabetic group the blood cholesterol level was 126.37 mg/dl. After induction of diabetic in the rats and treated with standard drug glyclazide and observed that the blood cholesterol level was reduce from 125.44 to 67.72 mg/dl. It was also observed that the various levels of cow urine  powder group (100, 200 and 400 mg) reduced significantly (p<0.01) from 134.23 to 95.55, from 132.88 to 93.70 and from 133.71 to 90.82 mg/dl respectively. In other hand LDL level for normal control group was 28.11 mg/dl and in diabetic group the blood LDL level was 86.87 mg/dl and in glyclazide group the blood LDL level was reduced from 83.73 to 16.17 mg/dl. Whereas in various levels of cow urine powder groups (100, 200 and 400 mg) reduced  significantly (p<0.01) from 99.35 to 51.15, from 98.43 to 47.22 and from 98.53 to 40.85 mg/dl respectively. HDL level was also estimated for normal control group was 53.53 mg/dl and in diabetic group the blood HDL level was 39.50 mg/dl. In glyclazide group the blood HDL level was increased from 41.75 to 51.55 mg/dl. Whereas in cow urine powder groups (100, 200 and 400 mg) increased significantly (p<0.01) from 34.90 to 44.40, from 34.42 to 46.47 and from 35.25 to 49.97 mg/dl respectively. Triglyceride level also measured for normal control group was 77.21 mg/dl and in diabetic group the blood triglyceride level was 188.67 mg/dl. Standard drug glyclazide group content the triglyceride level and it was reduced from 187.96 to 95.35 mg/dl. Other groups such as various levels of cow urine  powder groups (100, 200 and 400 mg) reduced significantly (p<0.01) from 189.36 to 134.55, from 179.73 to 132.70 and from 188.56 to 126.47 mg/dl respectively. Data obtained during the experiment conducted for standard drug group and cow urine powder groups (100, 200 and 400 mg) were significantly higher than the studies conducted by Gururaja et al.  (2011) on antidiabetic potential of cow urine distillate and found that significant reduction of the elevated blood glucose, serum cholesterol and serum triglyceride levels when compared with the diabetic control. The diabetic rats treated with cow urine distillate also showed a significant increase in HDL levels when compared with the diabetic controls and revealed the presence of antioxidants and free radicals scavengers in cow urine which might be responsible for the observed antidiabetic effect. Fig. Effect on glucose and lipid profile levels of diabetic rat treated with various control measures during the experiment 050100150200250300350400450BLOOD GLUCOSECHOLESTEROLLDLHDLTRIGLYCERIDE
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