The Preliminary Study Of Inhibitory Effect Of AWRK6 On Insulin Resistance And Its Mechanism

Insulin Resistance refers to the reduction of response to cell effect to physiological concentration of insulin that happens in the target tissue in internal,which makes the body produce compensatory islet beta cells to enhance its secretion,and finally leading to the blood insulin content higher than the normal range.Insulin resistance(IR)and the impaired insulin secretory of ? cell are the two major factors in the in the initiation and development of the Type 2 Diabetes Mellitus.An increasing number of studies have shown that IR was earlier abnormal in the development of T2 MD.It plays an important role in the development of the Type 2 Diabetes Mellitus.Antimicrobial peptides(AMP)is a low molecular weight peptide produced by organism itself to resist pathogen,with the features of broad-spectrum antibacterial activity,low immunogenicity and strong thermal stability.AWRK6 is a kind of AMPs which we reformed the peptides as 6 bases replacement the Dybowskin-2CDYa,which is better than Dybowskin-2CDYa.The sequence of AWRK6 is SWVGKHG KKFGLKKHKKH,which showed strong activity of insulin-secretion.The earlier stage results of our laboratory indicate that the mechanism of AWRK6 is similar to Exednin4 in promoting insulin secretion.Both of them can promote insulin secretion and the proliferation of ? cell effectively.In internal,they have strong stability and the obvious hypoglycemic effect,which make them expect to become a new generation of highly efficient and safe in treatment of the Type 2 Diabetes Mellitus.In recent years,it has been reported that Exendin4 can relieve insulin resistance and enhance insulin sensitivity.To investigate whether AWRK6 can inhibit insulin resistance,we choose the HepG2 cell and AWRK6 as the experiment object,using high concentration of insulin inducing the cell to establish the cell model of insulin resistance.We used MTT to identify the best preliminary of AWRK6.The methods of Glucose Oxidizing Enzyme and Triglyceride Oxidizing Enzyme are used to check the changes of glucose consumption and the amount of TG synthesis after AWRK6 treatment.The results are regarded as the standard to preliminary determine whether AWRK6 has an effect on insulin resistance.The results showed that the glucose consumption increased significantly and the amount of TG that intracellular synthesis increased significantly as well.It is proved that AWRK6 can relieve insulin resistance effectively.In order to further identify the inhibitory effect of AWRK6 on insulin resistance in internal,we used the method that the STZ was injected in the abdominal cavity of mice to establish the insulin resistance mice model.Applying OGTT method to judge the tolerance of Glucose in mice.And combining with ELISA tested the levels of insulin,TG,TC in blood and the levels of TG,TC,Glycogen in liver and skeletal muscle.The results showed that comparing with the model group,the tolerance of Glucose of mice was increased and the insulin sensitivity also enhanced.And the level of insulin was decreased significantly.But the levels of TG,TC,Glycogen were all boosted up in liver and muscle.These results further identified the inhibitory effect of AWRK6 on insulin resistance in internal.In order to explore the mechanism of AWRK6 inhibition of insulin resistance,we extracted the total protein in muscle of mice.We respectively detected the expression of PKB,PI3 K,GLUT4 with the method of Western Blot.The results showed that the expression of PKB,PI3 K,GLUT4 were all up-regulated after treatment with AWRK6.This study laid a further basic theoretical and experimental foundation for exploring the AWRK6 as a new type therapeutic for IR.