AWRK6 Combined With Epigallocatechin Gallate Attenuates Abnormal Glucose And Lipid Metabolism And Alleviates Type 2 Diabetes In Mice

Purposes: Type 2 diabetes(T2DM)is a chronic metabolic disease that seriously threatens human health.At present,the main T2 DM treatment is to control the blood sugar by blocking the body’s intake of protein and carbohydrates through medicines thus stimulating insulin secretion in islet tissue,or to increase the receptor activity of insulin.However,most of the current diabetes medicines have shortcomings such as difficulty in maintaining the homeostasis of blood glucose,short drug effect,and induction of discomfort in the digestive tract.Therefore,the development of new diabetes medicines is still essential for diabetes treatment.AWRK6(SWVGKHGKKFGLKKHKKH)is a cationic peptide obtained from the transformation of the forest frog skin antibacterial peptide,which has low immunogenicity and good stability.Animal experiments confirmed that AWRK6,as an agonist of GLP-1 receptor,has a good effect on promoting insulin secretion.AWRK6 can effectively alleviate the high-level fasting and postprandial blood glucose in T2 DM mice,and reduce the concentration of hemoglobin in mice induced by STZ.These data makes it a new promising and effective medicine for the treatment of type 2 diabetes.Epigallocatechin gallate(EGCG)is the main active ingredient of green tea and has significant antioxidant property,which can eliminate reactive oxygen species(ROS)produced by the respiratory chain in high sugar environment.Studies at home and abroad have shown that taking EGCG or foods containing EGCG can lower the level of blood sugar and triglycerides.Moreover,it has been reported that EGCG combined with other medicines,such as Exendin-4 and saponin compounds,can reduce blood sugar and blood lipids in T2 DM mice and enhance the antioxidant capacity of mice.Compared with using single medicine,combining different medicines is better to improve obesity and the metabolism disorder of glucose and lipid in T2 DM mice.This study analyzed the effect of combined AWRK6 and EGCG on glucose and lipid metabolism in T2 DM mice,and provided new ideas for the treatment of diabetes.Methods: In this study,a T2 DM mouse model was constructed by high-fat diet and STZ injection.The mice were divided into three groups and treated with AWRK6,EGCG and combined AWRK6 and EGCG,respectively.The combination group was intraperitoneally(i.p.)injected with AWRK6 and intragastrically(i.g.)administrated with EGCG.Normal and diabetes model mice were i.p.injected and i.g.administered with saline as control groups.Four weeks after treatment,the body weight,oral glucose tolerance,serum insulin and glycated hemoglobin were observed by immunohistochemistry.The effects of AWRK6 and EGCG combined therapy on type2 diabetes in mice were investigated.TC,TG,HDL-C and LDL-C content,calculate liver index,analyze the regulation of AWRK6,EGCG combined administration on lipid metabolism in type 2 diabetic mice;further use ELISA to detect serum corticosterone and 11β-hydroxyl The content of steroid dehydrogenase 1(11β-HSD1)was analyzed.The levels of total superoxide dismutase and malondialdehyde in mouse serum were analyzed,and the PI3K-AKT signaling pathway protein in mouse liver tissue was detected by western-blot technique.Expression,analysis of the possible mechanism of AWRK6 combined with EGCG in regulating glucose and lipid metabolism in type 2 diabetic mice.Results:(1)AWRK6,EGCG and their combination can reduce the body weight,blood glucose and serum insulin levels in type 2 diabetic mice.Compared with the T2 DM model control group,the serum insulin content of the mice in the co-administered group was significantly decreased(p<0.05),and the blood glucose was significantly decreased(p<0.01).Compared with the model group mice,the islet cells of the combined administration group were arranged neatly,with a large number of islets and a rounded shape.(2)AWRK6 and EGCG combined treatment can significantly reduce serum cholesterol and low-density lipoprotein cholesterol(P<0.05),increase serum high-density lipoprotein cholesterol(p<0.01),and decrease small in type 2 diabetic mice.Rat liver index(p<0.01).(3)AWRK6,EGCG and their combination can significantly increase the serum T-SOD content and decrease the serum MDA content in T2 DM mice.Compared withthe model control mice,the serum T-SOD content of the mice in the co-administered group was extremely significantly increased(p<0.01),and the serum MDA content was significantly decreased(p<0.01).The combination of AWRK6 and EGCG reduced serum corticosterone and 11β-HSD1 levels in type 2 diabetic mice,which was significantly different from the model control group(p<0.05).At the same time,AWRK6 combined with EGCG intervention can significantly increase the expression of p-PI3 K protein in liver tissue of type 2 diabetic mice(p<0.05),and significantly increase the expression of GLUT2 and p-AKT protein(p<0.01).Conclusions: AWRK6 combined with EGCG can reduce body weight and blood glucose in type 2 diabetic mice,protect islets from damage,reduce liver index,and improve dyslipidemia.AWRK6 and EGCG can improve the regulation of glucose and lipid metabolism and inhibit type 2 diabetes in mice by inhibiting oxidative stress,regulating hormone levels related to glycolipid metabolism,and activating PI3K-AKT signaling pathway.