摘　要：为探讨微小核糖核酸-133a（miR-133a）低表达对高糖诱导足细胞炎症凋亡的改善作用及相关机制，体外培养人肾小球足细胞（HGPC），并将其分为对照组（Con，不处理）、HG组（30 mmol/L D-葡萄糖）、inhibitor NC组（HG组+转染inhibitor NC）、miR-133a inhibitor组（HG组+转染miR-133a inhibitor）。转染24 h后，实时荧光定量逆转录PCR（RT-qPCR）检验转染效率。转染成功后，后续试验分为Con组、HG组、inhibitor NC组、miR-133a inhibitor组，miR-133a inhibitor+磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（AKT）通路抑制剂组（miR-133a inhibitor组+10 μmol/L LY294002），miR-133a inhibitor+PI3K/AKT通路激活剂组［miR-133a inhibitor组+50 mg/mL胰岛素样生长因子I（IGF-I）］。分别用CCK-8法、Hoechst 33258染色法、酶联免疫吸附试验（ELISA）、蛋白质印迹（Western blot）法检测HGPC细胞活力、凋亡率、肿瘤坏死因子-α（TNF-α）、白介素-6（IL-6）、PI3K、磷酸化（p）-PI3K、AKT、p-AKT的表达量。miR-133a转染成功。HG组和inhibitor NC组细胞活力、p-PI3K、p-AKT蛋白表达量低于Con组（P<0.05），细胞凋亡率、TNF-α、IL-6水平高于Con组（P<0.05）。miR-133a inhibitor组凋亡率、TNF-α、IL-6水平较inhibitor NC组低（P<0.05），p-PI3K、p-AKT蛋白表达升高（P<0.05）。LY294002减弱了miR-133a inhibitor的作用，IGF-I加强了miR-133a inhibitor的作用。miR-133a低表达可调控PI3K/AKT通路，抑制HG诱导HGPC细胞的炎症因子的释放和凋亡。本研究能够为糖尿病肾病相关疾病的研究提供新的方向。
关键词：糖尿病肾病；微小核糖核酸-133a；HGPC细胞；高糖；炎症
中图分类号：R364.5　　                  文献标志码：A　　            DOI：10.3969/j.issn.1007-7146.2025.03.009

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）