Metformin Regulating miR-34a Pathway to Inhibit Egr1 in Rat Mesangial Cells Cultured with High Glucose

Figure 6

SIRT1 gene silencing suppresses SIRT1 protein (by western blotting) expression in RMCs (a). Transfecting with SIRT1-siRNA obviously decreases phosphorylated AMPKα and increases Egr1 expression (by western blotting) though miR-34a inhibitor in high glucose-cultured RMCs (b). RMCs were transfected with SIRT1-siRNA (S-siRNA), nonspecific control (N-siRNA), or miR-34a inhibitor (IN) for 6 hours and further cultured with normal glucose (NG, 5.5 mmol/L glucose) or high glucose (HG, 30 mmol/L glucose) media for 24 hours. versus RMCs cultured with normal glucose media. ^#, ^## versus RMCs cultured with high glucose media. ^¢ versus RMCs transfected with miR-34a inhibitor. All results represent means  SD obtained from three independent experiments in triplicate and normalized to GAPDH. SIRT1: sirtuin 1; Egr1: early growth response factor 1; p-AMPKα: phosphorylated adenosine monophosphate-activated protein kinase α; AMPKα: adenosine monophosphate-activated protein kinase α; GAPDH: glyceraldehyde-3-phosphate dehydrogenase.

(a)

(b)