论文部分内容阅读
目的:探讨去甲斑蝥素对体外人源性肾小管细胞株(HK-2)上皮间质转分化以及转录因子Snail1的影响。方法:常规培养HK-2细胞,分为对照组、TGF-β1组、去甲斑蝥素组(NCTD组)。对照组为无血清DMEM培养,TGF-β1组为TGF-β15ng/ml诱导,NCTD组为不同浓度NCTD(0.5、1.0、2.5μg/ml)与TGF-β1(5ng/ml)共同作用,各组作用时间48h。采用RT-PCR、Western blot分别检测α-SMA、E-cadherin与Snail1表达水平的变化。结果:与对照组相比,TGF-β1组α-SMA mRNA和蛋白表达上调(P<0.05),E-cadherin mRNA和蛋白表达下调(P<0.05);与TGF-β1组相比,NCTD干预组α-SMA mRNA和蛋白表达下调(P<0.05),而E-cadherin mRNA和蛋白表达上调(P<0.05),且均呈剂量依赖性。与对照组相比,TGF-β1组Snail1 mRNA和蛋白表达上调(P<0.05);与TGF-β1组比较,NCTD干预组Snail1 mRNA和蛋白表达下调(P<0.05),具有一定的剂量依赖关系。结论:去甲斑蝥素具有抑制肾小管上皮细胞EMT的作用,该作用可能与下调转录因子Snail1的表达有关。
Objective: To investigate the effect of norcantharidin on the transdifferentiation of epithelial mesenchyme and the transcription factor Snail1 in human renal tubular cell line (HK-2) in vitro. Methods: HK-2 cells were cultured routinely and divided into control group, TGF-β1 group and norcantharidin group (NCTD group). The control group was induced by serum-free DMEM, the TGF-β1 group was induced by TGF-β15ng / ml, the NCTD group was treated by different concentrations of NCTD (0.5,1.0,2.5μg / ml) and TGF- The role of time 48h. The expression of α-SMA, E-cadherin and Snail1 were detected by RT-PCR and Western blot respectively. Results: Compared with the control group, the expression of α-SMA mRNA and protein in TGF-β1 group was up-regulated and the expression of E-cadherin mRNA and protein were down-regulated (P <0.05) The mRNA and protein expression of α-SMA were down-regulated (P <0.05), while the expression of E-cadherin mRNA and protein was up-regulated (P <0.05), both of which were dose-dependent. Compared with the control group, the expression of Snail1 mRNA and protein in TGF-β1 group was up-regulated (P <0.05). Compared with TGF-β1 group, the expression of Snail1 mRNA and protein in NCTD intervention group was down-regulated . Conclusion: Norcantharidin can inhibit the EMT of renal tubular epithelial cells, which may be related to the down-regulation of Snail1.