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目的探讨微小RNA(microRNA,miRNA)-451对肾小球系膜细胞增殖的抑制作用及其机制。方法在Lipofectamine 2000介导下,将miR-451模拟物(miR-451 mimics)及mimics对照分别转染高糖培养的肾小球系膜细胞,另设高糖未转染组和低糖未转染组,实时荧光RT-PCR法检测各组细胞中miR-451的表达水平;MTT法检测miR-451对小鼠肾小球系膜细胞增殖的影响;Western blot法检测靶蛋白Ywhaz、p-p38 MAPK和p-MKK3的表达水平。结果 miR-451组细胞中miR-451的表达水平较mimics对照组明显升高(P<0.01);miR-451组第2、3、4天的细胞增殖能力明显低于高糖未转染组(P均<0.05),第3、4天明显低于mimics对照组(P均<0.05);与mimics对照组和高糖未转染组比较,miR-451组细胞中Ywhaz、p-p38 MAPK和p-MKK3的表达水平均明显下降(P均<0.05)。结论 miR-451可抑制Ywhaz蛋白的表达,降低p38 MAPK信号途径中p-p38 MAPK和p-MKK3蛋白的表达,从而降低高糖诱导的肾小球系膜的增殖。本实验为研究糖尿病肾病(diabetic nephropathy,DN)的发病机制提供了新的思路。
Objective To investigate the inhibitory effect of microRNA (miRNA) -451 on mesangial cell proliferation and its mechanism. METHODS: miR-451 mimics and mimics were transfected into mesangial cells cultured in high glucose medium under the control of Lipofectamine 2000, respectively. High-glucose non-transfected and untransfected The expression of miR-451 in each group was detected by real-time fluorescent RT-PCR. The effect of miR-451 on the proliferation of mesangial cells was detected by MTT assay. The expression of target protein Ywhaz, p-p38 MAPK and p-MKK3 expression levels. Results The miR-451 expression in miR-451 group was significantly higher than that in mimics control group (P <0.01). The proliferation of miR-451 group on day 2,3 and 4 was significantly lower than that of the untreated group (P <0.05), and were significantly lower than those of mimics control group on the 3rd and 4th day (all P <0.05). Compared with mimics control group and high glucose untransfected group, the expression of Ywhaz, p-p38 MAPK And p-MKK3 expression were significantly decreased (P all <0.05). Conclusion miR-451 can inhibit the expression of Ywhaz protein and down-regulate the expressions of p-p38 MAPK and p-MKK3 in the p38 MAPK signaling pathway, thereby reducing the high glucose-induced mesangial proliferation. This experiment provides a new idea for studying the pathogenesis of diabetic nephropathy (DN).