The Effect And Mechanism Of AWRK6 On Improving Insulin Resistance

Insulin resistance(IR)refers to the sensitivity of insulin production and the relative decrease in reactivity.The body maintains a normal concentration of glucose and compensatoryly secretes insulin,eventually leading to hyperinsulinemia.IR is the core pathological mechanism that induces the development of type 2 diabetes(T2DM).Some studies have shown that IR is an early physiological abnormality of T2 DM,which plays an irreplaceable role in the formation of T2 DM.AWRK6(SWVGKHGKKFGLKKHKKH)is a novel bioactive peptide,which is a novel antibacterial peptide optimized by replacing 6 bases with the antibacterial peptide Dybowskin-2CDYa(SAVGRHGRRFGLRKH RKH).Previous studies have shown that AWRK6 can promote the secretion of insulin by MIN6 cells,which is a novel GLP-1 receptor agonist.It has significant therapeutic effects on T2 DM mice,and its mechanism of action is similar to that of Exendin-4.In this study,HepG2 cells were treated with high concentration of insulin in vitro,an IR cell model was established,and the optimal concentration of AWRK6 and the positive drug Exendin-4 was screened.The glucose consumption of HepG2 cells in blank group,IR group,Exendin-4 positive control group and AWRK6 experimental group was detected by GOD-POD method,and the improvement effect of AWRK6 on IR was preliminarily investigated.At the same time,we used male Kunming mice,randomly divided into blank group(15)and HFD model group(45).The blank group was given feed to standard normal mice,and the model group was given high-fat diet for 12 weeks to establish IR.Mouse model.The insulin resistance index(HOMA-IR)and insulin sensitivity index(ISI)assessed by the steady-state model were calculated as a measure of modeling by testing fasting blood glucose(FBG)and fasting serum insulin(FINS).After successful modeling,the model mice were randomly divided into three groups:HFD model group,AWRK6 experimental group and Exendin-4 positive control group.After 4 weeks of drug intervention,the mice were subjected to oral glucose tolerance test(OGTT)and insulin tolerance test.(IPITT),using a chemical test kit to measure fasting serum insulin(FINS),serum glucagon(FGC),glycated hemoglobin(HbA1c),total cholesterol(T-CHO),triglyceride(TG),high density lipid Protein(HDL-C),low-density lipoprotein(LDL-C),hepatic glycogen and muscle glycogen wereobserved by HE staining and oil red O staining to observe the pathological changes of liver tissue and evaluate the effect of AWRK6 on IR.The effects of AWRK6 on the expression of downstream insulin-related proteins PI3 K,P-PI3 K,AKT,P-AKT and GLUT2 were studied in vitro and in vivo by immunoblotting.The mechanism of AWRK6 on IR improvement was analyzed.The results showed that high concentration of insulin interfered with HepG2 cells,and the supernatant glucose content increased significantly,resulting in IR.The intervention of AWRK6 significantly increased the glucose consumption of IR model cells.After 12 weeks of high-fat diet,the HFD model group mice and blank group Compared with the FGB,FINS,HOMA-IR and other waters with extremely significant increase,and HOMA-IR ? 2.7,the IR model was established successfully;compared with the model,AWRK6 intervention began 1 week,the mice lost weight,fasting Blood glucose levels decreased;AWRK6 significantly attenuated glucose tolerance and insulin tolerance after 4 weeks of drug intervention,while significantly reducing FGB,FINS,HOMA-IR,ISI levels,and HbA1 c,TG,T-CHO,LDL-C levels,and significantly increased The levels of FGC,HDL-C,hepatic glycogen and muscle glycogen;HE staining and oil red O staining showed that liver tissue sections showed that AWRK6 can significantly improve liver lipid degeneration and lipid deposition;AWRK6 can significantly increase cell level And the expression of P-PI3 K,P-AKT,GLUT2 in the tissue level,promote glucose uptake and improve IR.The results of this study indicate that AWRK6 has an inhibitory effect on insulin resistance in vitro and in vivo,and up-regulates PI3K/P-PI3K/AKT/P-AKT/GLUT2 signaling pathway to promote glucose utilization and improve insulin resistance,laying a foundation for further clinical drug development.The experimental basis and theoretical basis.