Objective To investigate the regulation of insulin VGLUT2 gene expression in pancreatic β cells and its exactly mechanism.Methods The rat pancreatic β cell line RIN-5F was treated by different insulin concentrations and different siRNA.The changes of VGLUT2 mRNA and protein expression levels were detected by adopting real-time qPCR and Western blot.Results In-sulin with a concentration of 100,200 nmol/L significantly inhibited the expressions of VGLUT2 mRNA and protein in RIN-5RF cells(P<0.05),moreover the inhibiting effect was most significant at 100 nmol/L.After 100 nmol/L insulin treatment,the expres-sions of VGLUT2 mRNA and protein at 12,18,24 h were significantly inhibited compared with that at 0 h(P<0.05).Compared with the Blank group,Lip2000 group and Control-siRNA group,after interfering RIN-5F by using IR-siRNA,IRS1-siRNA and IRS2-siRNA,the inhibition situation of VGLUT2 mRNA and protein expressions by 100 nmol/L insulin in each group was signifi-cantly recovered(P<0.05).Conclusion Insulin at low concentration could inhibit VGLUT 2 gene expression in rat pancreatic β cell line RIN-5F.