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探讨Micro RNA-217(Mir-217)、低氧诱导因子1α(HIF-1α)及沉默信息调节因子1(Sirt1)在大鼠肾小球系膜细胞炎性反应及纤维化发生过程中的作用。用高糖培养的大鼠肾小球系膜细胞,采用RT-PCR技术检测Mir-217、HIF-1αm RNA及Sirt1的m RNA表达水平,采用Western-blotting技术检测HIF-1α、Sirt1、纤连蛋白(FN)、内皮素1(ET-1)、结缔组织生长因子(CTGF)表达水平,采用ELISA法检测血管内皮生长因子(VEGF)及转化生长因子-β1(TGF-β1)表达水平。结果显示,高糖刺激可下调大鼠肾小球系膜细胞中的Sirt1的表达水平,上调Mir-217、HIF-1α、FN、ET-1、CTGF、VEGF及TGF-β1的表达水平,差异具有统计学意义(p<0.05);而采用Sirt1特异性激活剂白藜芦醇预处理或Mir-217基因沉默可上调Sirt1表达水平,下调Mir-217、HIF-1α、FN、ET-1、CTGF、VEGF及TGF-β1的表达水平,差异具有统计学意义(p<0.05);因此,本研究认为Mir-217可以通过调节Sirt1/HIF-1α通路来调控肾小球系膜细胞炎性反应及纤维化过程。
To investigate the role of micro RNA-217 (Mir-217), hypoxia inducible factor 1α (HIF-1α) and silencing information-regulated factor 1 (Sirt1) in inflammatory reaction and fibrosis in rat glomerular mesangial cells . The mesangial cells were cultured with high glucose. The m RNA expression of Mir-217, HIF-1αmRNA and Sirt1 was detected by RT-PCR. Western blotting was used to detect the expression of HIF-1α, Sirt1, The expression of vascular endothelial growth factor (VEGF) and transforming growth factor-β1 (TGF-β1) were detected by enzyme linked immunosorbent assay (ELISA). The expressions of FN, ET-1 and CTGF were detected. The results showed that high glucose stimulation could down-regulate the expression of Sirt1 in rat mesangial cells and increase the expression of Mir-217, HIF-1α, FN, ET-1, CTGF, VEGF and TGF-β1 (P <0.05). Sirt1-specific activator resveratrol pretreatment or Mir-217 gene silencing could up-regulate Sirt1 expression and down-regulate Mir-217, HIF-1α, FN and ET- CTGF, VEGF and TGF-β1, the difference was statistically significant (p <0.05); therefore, this study suggests that Mir-217 can regulate the Sirt1 / HIF-1α pathway to regulate the inflammatory response of mesangial cells And fibrosis process.