泽泻醇B抑制C3a介导的人肾小管上皮细胞间充质转分化的实验研究

Alisol B Inhibited Complement 3a-induced Human Renal Tubular Epithelial to Mesenchymal Transition

目的探讨泽泻醇B能否抑制C3a介导的肾小管上皮细胞间充质转分化(epithelial-mesenchymal transition,EMT)。方法将体外培养的人肾小管上皮细胞(HK-2)分别用5ng/mL转化生长因子β(transfor-ming growth factor-β,TGF-β)、0.1μmolC3a和0.1μmolC3a加10μmol泽泻醇B进行干预。分别采用RT-PCR、Western blot和细胞免疫荧光法检测HK-2细胞C3、α-平滑肌肌动蛋白(α-SMA)和上皮钙黏蛋白(E-cadherin)mRNA及蛋白表达。结果经C3a刺激后,HK-2细胞C3mRNA和蛋白表达明显增加(P<0.01),α-SMA mRNA及蛋白表达明显增加(P<0.01),E-cadherin mRNA及蛋白表达明显减少(P<0.01)。与经外源性C3a干预组比较,经泽泻醇B干预后,HK-2细胞α-SMA mRNA及蛋白表达明显减少(P<0.01),E-cad-herin mRNA及蛋白表达明显增加(P<0.05)。结论外源性C3a能诱导肾小管上皮细胞发生EMT,泽泻醇B可抑制C3a诱导的EMT。

Objective To study whether alisol B could inhibit complement 3a （C3a） induced renal tubular epithelial-mesenchymal transition （EMT）. Methods The in vitro cultured human renal tubular epithelial HK-2 cells were intervened with 5 ng/mL transforming growth factor-[3 （TGF-y）, 0. 1 lamol C3a, and 0. 1 12mol C3a ＋ 10 μmol alisol B, respectively. The mRNA and protein expressions of a-SMA, E-cadherin, and C3 were detected using RT-PCR, Western blot, and immunofluorescence, respectively. Results The mRNA and protein expres- sions of C3 in HK-2 cells were up-regulated after intervention of C3a （P〈0. 01 ）, the mRNA and protein expres- sions of a-SMA in HK-2 cells were obviously enhanced （ P 〈0.01 ）, the mRNA and protein expressions of E-cad- herin obviously decreased （ P 〈 0.01 ）. When compared with the group intervened by exogenous C3a, after inter- vention of alisol B, the mRNA and protein expressions of a-SMA in HK-2 cells were obviously reduced （ P 〈 0.01 ）, the mRNA and protein expressions of E-cadherin obviously increased （ P 〈 0.05）. Conclusions Exoge- nous C3a could induce renal tubular EMT. Alisol B was capable of suppressing C3a induced EMT.