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Abstract Abstract: To investigate the effect of low expression of microRNAs-133a (miR-133a) on the apoptosis of podocytes induced by high glucose and its related mechanism, human glomerular podocyte (HGPC) cells were cultured in vitro and divided into control group (Con, no treatment), HG group (30 mmol/L D-glucose), inhibitor NC group (HG group+transfection inhibitor NC), miR-133a inhibitor group (HG group+ transfection miR-133a inhibitor). 24 h after transfection, real-time fluorescence quantitative PCR (RT-qPCR) was used to detect the transfection efficiency. Follow-up tests were performed after transfection success, which were divided into Con group, HG group, inhibitor NC group and miR-133a inhibitor group, miR-133a inhibitor+phosphatidylinositol 3-kinase (PI3K) / protein kinase B (AKT) pathway inhibitor group (miR-133a inhibitor+10 μmol/L LY294002), miR-133a inhibitor+PI3K/AKT pathway activator group [miR-133a inhibitor+50 mg/mL insulin-like growth factor (IGF-I)]. Live cell counting method (CCK-8) and Hoechst 33258 staining were used respectively, enzyme-linked immunosorbent assay (ELISA) and Western blot were used to detect HGPC cell viability, apoptosis rate, tumor necrosis factor (TNF) -α, interleukin-6 (IL-6) levels, PI3K, phosphorylation (p) -PI3K, AKT and p-AKT expression levels. miR-133a was transfected successfully. The cell viability, p-PI3K and p-AKT protein expression of HG group and inhibitor NC group were lower than those of Con group (P<0.05), the apoptosis rate, TNF-α and IL-6 levels were higher than those of Con group (P<0.05). The apoptosis rate, TNF-α and IL-6 levels of miR-133a inhibitor group were lower than those of miR-133a inhibitor group (P<0.05), while the protein expressions of p-PI3K and p-AKT increased (P<0.05). LY294002 attenuated the action of miR-133a inhibitor and IGF-I enhanced the action of miR-133a inhibitor. The low expression of miR-133a can regulate the PI3K/AKT pathway to inhibit the release of inflammatory factors and apoptosis of HG-induced HGPC cells, this research could provide a new direction for the study of diseases related to diabetic nephropathy.
Key words: diabetic nephropathy; miR-133a; HGPC cells; high glucose; inflammation
（Acta Laser Biology Sinica, 2025, 34(3): 267-273）
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