Ssc-miR-146b Inhibits Glucose Uptakeby Targeting IRS1 In Porcine Adipocyte

Adipose is the largest energy reserve system in both animal and human,andit is an important place for glycolipid metabolism.The regulation of blood glucose is mainly regulated by insulin / IR / IRS1 / PI3 K / GLUT4 signaling pathway.micro RNAs(mi RN As)are involved in many physiological processes of adipocytes,including lipid metabolism and cell differentiation.O n the basis of the previous study,ssc-mi R-146 b reduced the glucose uptake in porcine adipocytes.The predicted target gene of ssc-mi R-146 b was conducted by bioinformatics method.Western blot,luciferase reporter system were used for the evaluation of the regulating function of ssc-mi R-146 b.Results are shown as follows:1.KEGG analysis revealed that ssc-mi R-146 b was involved in the signal transduction pathway of adipocytokines.Both GLUT4 and IRS1 were the predicted tar gets of ssc-mi R-146 b.2.After transfection of ssc-mi R-146 b mimics / NC / inhibitor / i NC,western blot showed that ssc-mi R-146 b decreased both the GLUT4 protein and IRS1 protein but the result of pmir GLO dual luciferase assay claimed that GLUT4 was not the target for ssc-mi R-146 b,IRS1 instead.3.Three kinds of mutants of mi R-146 b were constructed,pmir GLO dual luciferase assay showed that mutant 1 and mutant 2 still targeted 3'-UTR of IRS1 while mutant 3 failed to target IRS1,which indicatedthe 18 th and 21 st base of ssc-mi R-146 b played a significant role in ssc-mi R-146 b targets.In conclusion,this study indicated that ssc-mi R-146 b could reduce the uptake ofglucose in porcine adipocytes via suppressing the expression of GLUT4 by targeting IRS1.