丹参酚酸B盐对TGF-β1诱导的HK-2上皮-间充质转分化的影响

Impact of salvianolic acid-B on TGF-β1-induced HK-2 epithelial-mese nchymal transition

目的:研究丹参酚酸B盐(salvianolic-acid B,SA-B)对转化生长因子β1(transforming growth factorβ1,TGF-β1)诱导的人肾小管上皮细胞HK2(human human renal proximal tubular cells)转分化的影响。方法:复苏并传代培养HK2细胞,应用不同剂量的TGF-β1或同一剂量作用不同时间诱导HK2细胞发生上皮-间充质转分化(epithelia-mesenchymal transition,EMT),同时给予骨形成蛋白7(BMP-7,阳性对照)(50μg.L-1)和SA-B(1×10-5mol.L-1)干预观察二者对EMT的影响。应用Western印迹法和免疫荧光法检测HK2细胞E-钙黏蛋白(E-cadherin),α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的表达。结果:BMP-7显著抑制了TGF-β1诱导的E-cadherin下调(P<0.05)和α-SMA上调(P<0.05);而SA-B显著抑制TGF-β1诱导的α-SMA表达增加(P<0.05),未见其抑制TGF-β1诱导的E-钙黏蛋白下调。结论:SA-B和BMP-7能够抑制TGF-β1诱导的HK2上皮-间充质转分化,其共同的作用是抑制了TGF-β1诱导的α-SMA上调,而SA-B对E-钙黏蛋白的调控作用有待进一步的研究。

Objective： To investigate the effect of salvianolic-acid B （SA-B） on epithelia-mesenchymal transition in human renal proximal tubular cells （ HK2 ）, induced by transforming growth factor β1 （ TGF-β1 ）. Method： Epithelia-mesenchymal transition （EMT） was induced with TGF-fll in HK2 cultured in vitro. Different concentrations （2, 5, 10, 20 μg . L-1 ） and stimulant periods （12, 24, ,18 h ） were tried to find the perfect condition for EMT. At the same time bone morphogenetic protein-7 （BMP-7, positive control） and the SA-B intervention were given to observe their effect on EMT. Western blot and immunofluorescent microscopy were used to analyze the expression of E-cadherin and α-smooth muscle actin （α-SMA） in HK2. Result： BMP-7 significantly inhibited the down-regulation of E-cadherin and the up-regulation of a-SMA induced by TGF-β1 （ P 〈 0. 05 ）, and SA-B significantly inhibited the up-regulation of a-SMA expression induced by TGF-fll （ P 〈 0.05 ）, but not the down-regulation of E-cadherin induced by TGF-fll. Conclusion ： SA-B and BMP-7 can inhibit TGF-β1-induced EMT in HK2. Their common role is to inhibit the up-regulation of α-SMA, and the effect of SA-B on the regulation of E-cadherin needs further study to be confirmed.