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目的:研究丹酚酸B逆转转化生长因子(TGF)-β1诱导的肾小管上皮细胞-间充质细胞转分化时microRNA(miRNA)表达谱的变化。方法:将体外培养的人近端肾小管上皮细胞系(HK-2)细胞分为3组:①对照组:未加入丹酚酸B或者TGF-β1;②TGF-β1组:在细胞培养基中加入TGF-β1(浓度为5ng/mL);③丹酚酸B逆转组:在细胞培养基中先TGF-β1(浓度为5ng/mL),48h后HK-2细胞成功诱导EMT后形成的间充质细胞,再更换为TGF-β1(浓度为5ng/mL)和丹酚酸B(50μmol/L)继续培养72h,采用倒置相差显微镜观察细胞形态学变化;免疫细胞化学染色检测细胞E-cadherin的表达情况;基因芯片检测各组细胞miRNA的表达变化。结果:①丹酚酸B具有逆转HK-2细胞EMT的作用;②与正常HK-2细胞比较,TGF-β1组有25种miRNA的表达出现显著改变,其中15种表达下调2倍以上,10种表达上调2倍以上。③与TGF-β1组比较,丹酚酸B逆转组,有44种miRNA的表达出现显著改变,其中24种表达下调2倍以上,20种表达上调2倍以上。④有10种miRNA表达在两组同时出现显著改变,其中8种在TGF-β1组明显下调,而在丹酚酸B逆转组明显上调,1种在TGF-β1组明显上调,而在丹酚酸B逆转组则明显下调。结论:丹酚酸B具有阻止慢性肾脏疾病进行性发展的潜能,而这一作用与其能有效调控miRNA的表达有关。
AIM: To investigate the effect of salvianolic acid B on reversing the expression of microRNA (miRNA) in renal tubular epithelial-mesenchymal cells induced by transforming growth factor-β1 (TGF-β1). Methods: Human proximal tubular epithelial cell line (HK-2) cells cultured in vitro were divided into three groups: ① control group: no salvianolic acid B or TGF-β1; ②TGF-β1 group: in the cell culture medium ③ TGF-β1 (5ng / mL) was added to TGF-β1 at a concentration of 5ng / mL. ③ Reverse transfection group of salvianolic acid B: TGF- The cells were treated with TGF-β1 (5ng / mL) and salvianolic acid B (50μmol / L) for 72h. The morphological changes of cells were observed by inverted phase contrast microscope. The expressions of E-cadherin The expression of miRNA in each group was detected by microarray. RESULTS: Salvianolic acid B reversed the effect of EMT in HK-2 cells. (2) Compared with normal HK-2 cells, the expressions of 25 miRNAs in TGF-β1 group were significantly changed, of which 15 were down-regulated more than 2 times and 10 The species is upregulated more than 2 times. ③Compared with TGF-β1 group, the expression of 44 miRNAs changed significantly in salvianolic acid B reversed group, of which 24 were down-regulated more than 2 times and 20 more than 2 times. ④ There were 10 kinds of miRNA expression in the two groups at the same time significant changes, of which 8 were significantly down-regulated in the TGF-β1 group, while in the salvianolic acid B reversed group was significantly increased, a was significantly increased in the TGF-β1 group, Acid B reversed the group was significantly reduced. Conclusion: Salvianolic acid B has the potential to prevent the progressive development of chronic kidney disease, which is related to its ability to regulate miRNA expression.