【摘要】 目的：探讨类甲基化转移酶3(METTL3)/miR-126对高糖（HG）诱导的大鼠肾小管上皮细胞（NRK-52E）增殖的影响及其作用机制。方法：将NRK-52E细胞分为对照组（5.5 mmol/L葡萄糖）、HG组（35 mmol/L葡萄糖）、HG+siRNA-NC组和HG+METTL3 siRNA组。采用MTT法检测细胞增殖，流式细胞仪检测细胞周期，酶联免疫吸附试验（ELISA）法检测细胞上清液中转化生长因子（TGF）-β1蛋白含量，RT-qPCR法检测miR-126和METTL3基因表达，western blotting检测METTL3和smad2蛋白表达，RNA免疫共沉淀技术（RIP）检测METTL3与miR-126的结合情况。结果：与对照组比较，HG组细胞增殖率和G1期细胞所占百分比降低，G2期细胞所占百分比及METTL3、TGF-β1、smad2、miR-126表达升高（P<0.05）；与HG组比较，HG+METTL3 siRNA组细胞增殖率升高，G1期细胞所占百分比降低，S期细胞所占百分比升高，METTL3、TGF-β1、smad2、miR-126表达降低（均P<0.05）。HG组与HG+siRNA-NC组各指标比较，差异均无统计学意义（P>0.05）。RIP实验结果显示，METTL3siRNA可降低成熟miR-126的含量（P<0.05）。结论：METTL3在HG诱导的肾小管上皮细胞中的表达上调，抑制细胞增殖，其可能与甲基化修饰miR-126并激活TGF-β1/smad2信号通路有关。更多还原

【Abstract】 Objective: To investigate the effects of methyltransferase-like 3 (METTL3)/miR-126 on hyperglycemia (HG)-induced proliferation of normal rat kidney-52E (NRK-52E) and its mechanism. Methods: NRK-52E cells were divided into control group (5.5 mmol/L glucose), HG group (35 mmol/L glucose), HG + siRNA- NC group and HG+METTL3 siRNA group. Cell proliferation was detected by MTT, cell cycle was detected by flow cytometry, transforming growth factor (TGF)- β1 protein content in cell supernatant was detected by enzymelinked immunosorbent assay (ELISA), gene expression of miR- 126 and METTL3 was detected by RT- qPCR METTL3 and smad2 protein expression was detected by western blotting, and the binding of METTL3 to miR-126 was detected by RNA immunoprecipitation (RIP). Results: Compared with the control group, the cell proliferation rate and the percentage of G1- phase cells were decreased in the HG group, and the percentage of G2-phase cells and the expression of METTL3, TGF-β1, smad2 and miR-126 were increased in the HG+METTL3siRNA group (P<0.05). Compared with the HG group, the cell proliferation rate was increased, the percentage of G1-phase cells was decreased, the percentage of S-phase cells was increased, and the expression of METTL3TGF-β1, smad2, and miR-126 was decreased in the HG + METTL3 siRNA group (P<0.05). There was no statistically significant difference between the HG group and the HG + siRNA-NC group in any index (P>0.05). The results of RIP experiment showed that METTL3siRNA could reduce the content of mature miR-126(P<0.05). Conclusion: METTL3 expression is upregulated in HG-induced renal tubular epithelial cells and inhibit cell proliferation, which may be related to the methylation modification of miR-126 and the activation of TGF-β1/smad2 signaling pathway.更多还原