Effect of recombinant human resistin on glucose metabolism by AMPK pathway in HepG2 cells / 实用医学杂志

ABSTRACT

Objective To investigate the effect of rh-resistin on glucose metabolism in HepG2 cells and to elucidate whether the underlying mechanisms are related to AMPK pathway. Methods Cells transfected with control siRNA or AMPKα2 siRNA were cultured in 6-well plates and then treated with 50 ng/mL rh-resistin for 24 hours , while untransfected cells were treated with or without 50 ng/mL rh-resistin on the same conditions , followed by serum-starving in glucose-free DMEM for 3 ~ 5 hours in the continued absence or presence of rh-re-sistin. Then the cells were treated with or without insulin for 2 hours. AMPKα2, G6Pase, PEPCK and Glut2 mRNA expression levels were determined by quantitative RT-PCR. The phosphorylation state of AMPK was deter-mined by Western blotting. Glycogen synthesis was measured by the incorporation of D-[U-14C] glucose to glycogen. Results Rh-resistin suppressed the AMPKα2, Glut2 mRNA expressions, and reduced the phosphory-lation level of AMPK and glycogen synthesis on both basal and insulin-stimulated conditions (P < 0.05), while it accelerated G6Pase and PEPCK mRNA expressions on the same conditions (P < 0.05). The mRNA expression levels of G6Pase , PEPCK , Glut2 and the phosphorylation level of AMPK and glycogen synthesis were signifi-cantly different between the rh-resistin group and the rh-resistin in conjunction with AMPKα2 siRNA-treated group. Conclusion Rh-resistin may affect glucose metabolism in HepG2 cell via AMPK pathway.