MiR-138-5p affects insulin resistance to regulate type 2 diabetes progression through inducing autophagy in HepG2 cells by regulating SIRT1

Abstract

Insulin resistance (IR) is considered as a major factor of type 2 diabetes (T2D), which is seriously detrimental to human health. In our present study, we found that the expression of miR-138-5p was increased in the insulin-resistant HepG2 cells induced by TNF-α. Therefore, we hypothesized that miR-138-5p might play a regulatory role in the IR. To examine this hypothesis, HepG2 cells were transfected with miR-138-5p inhibitor. Silencing of miR-138-5p increased glucose uptake and glycogen synthesis of TNF-α–stimulated HepG2 cells and decreased glucose concentration in medium, suggesting that downregulation of miR-138-5p suppressed IR in HepG2 cells. Besides that, we found that sirtuin 1 (SIRT1) was the target gene of the miR-138-5p. Moreover, co-transfection with SIRT1-siRNA and miR-138-5p inhibitor suppressed glucose uptake and glycogen synthesis of HepG2 cells compared with miR-138-5p inhibitor–transfected group, indicating that downregulation of SIRT1 weakened the inhibitory effect of miR-138-5p inhibitor on IR. In addition, overexpressed SIRT1 increased Beclin1, LC3 II/I level, and the number of GFP-LC3 dots and decreased p62 level, whereas downregulation of SIRT1 had the opposite effects. Our results demonstrated that overexpressed SIRT1 activated autophagy in HepG2 cells. Moreover, we observed that 3-methyladenine (an inhibitor of autophagy) treatment decreased the high glucose uptake and glycogen synthesis of miR-138-5p inhibitor–transfected HepG2 cells, suggesting that the inhibition of autophagy abolished the inhibitory effect of miR-138-5p inhibitor on IR in HepG2 cells. Taken together, this study suggested that miR-138-5p contributed to the TNF-α–induced IR, possibly through inducing autophagy in HepG2 cells by regulating SIRT1. MiR-138-5p might be a potential and promising target for the treatment of IR.