下调miR-433表达抑制高糖介导的肾纤维化机制研究

The suppression of miR-433 expression reduces renal fibrosis induced by high glucose

目的探讨下调表达miR-433抑制高糖刺激下肾纤维化的机制。方法实时荧光定量PCR(qRT-PCR)检测体外高糖刺激的大鼠原代肾小管上皮细胞(TECs)和肾小球系膜细胞(MMCs)中miR-433水平及纤维化指标:胶原Ⅰ、Ⅳ(Col-Ⅰ,Col-Ⅳ)、粘连蛋白(FN)和α平滑肌肌动蛋白(α-SMA)的表达。高糖刺激稳定表达miR-433上调及miR-433下调NRK52E和MMCs细胞系,从核酸和蛋白水平检测上述纤维化指标。结果高糖刺激的TECs和MMCs中miR-433、Col-Ⅰ、Col-Ⅳ、FN和α-SMA的表达升高(P<0.001)。高糖刺激下,上调miR-433表达的TECs和MMCs中miR-433和纤维化指标的表达明显升高(P<0.001),而下调miR-433表达TECs和MMCs中miR-433和纤维化指标的表达明显抑制(P<0.001)。结论高糖刺激下,大鼠TECs和MMCs miR-433和纤维化指标表达升高,下调miR-433表达可抑制高糖介导的肾纤维化;miR-433表达可作为糖尿病肾脏疾病潜在治疗靶点。

Objective To investigate the mechanism of miR-433 suppression in inhibiting renal fibrosis induced by high glucose.Methods Rat tubular epithelial cells(TECs)and glomerular mesangial cells(MMCs)were stimulated by high glucose.Expression of miR-433,Col-Ⅰ,Col-Ⅳ,FN andα-SMA were assessed by qRT-PCR and Western Blot.Results The expression of miR-433,Col-Ⅰ,Col-Ⅳ,FN andα-SMA in TECs and MMCs stimulated by high glucose was significantly increased(P<0.001).MiR-433 and fibrotic markers of TECs and MMCs were significantly increased when the expression of miR-433 was up-regulated(P<0.001).Conclusion High glucose stimulates rat TECs and MMCs to express miR-433 and fibrotic markers in time-and dose-dependent manners.Down-regulation of miR-433 suppresses renal fibrosis induced by high glucose.