止消通脉宁对TGF-β_1诱导的人肾小管上皮细胞CTGF、PAI-1、MMP-9 mRNA表达的影响

Effect of Zhixiaotongmaining on Expression of CTGF, PAI-1 and MMP-9 mRNA of Human Renal Tubular Epithelial Cell HK-2 Induced by TGF-β_1

目的观察止消通脉宁药物血清(以下简称"药物血清")清干预转化生长因子-β1(TGF-β1)诱导的人肾小管上皮细胞(HK-2)结缔组织生长因子(CTGF)、纤溶酶原激活物抑制物-1(PAI-1)、基质金属蛋白酶-9(MMP-9)mRNA的表达,探讨其在防治肾间质纤维化方面的作用。方法将HK-2细胞用含10%胎牛血清的DMEM/F12(1∶1)培养基培养。实验分为空白对照组、单纯TGF-β1诱导组(TGF-β110 ng/mL)、空白血清对照组(TGF-β110 ng/mL+10%空白血清)、干预1组(TGF-β110 ng/mL+10%低剂量药物血清)、干预2组(TGF-β110 ng/mL+10%中剂量药物血清)、干预3组(TGF-β110 ng/mL+10%高剂量药物血清)。药物干预24 h后,荧光定量PCR检测CTGF、PAI-1和MMP-9 mRNA的表达。结果 HK-2细胞经TGF-β1诱导后,CTGF、PAI-1 mRNA表达显著上升,MMP-9 mRNA表达减弱,与空白对照组比较差异有统计学意义(P<0.05)。经止消通脉宁药物血清干预后,CTGF、PAI-1 mRNA表达逐步下降,MMP-9 mRNA表达逐步上升,与单纯TGF-β1诱导组比较差异有统计学意义(P<0.05)。结论止消通脉宁在一定程度上能够抑制TGF-β1诱导的人肾小管上皮细胞纤维化,其机制可能与调节纤维化细胞因子CTGF、PAI-1和MMP-9的mRNA表达有关。

Objective To observe the effect of Zhixiaotongmaining on the expression of CTGF, PAl-1 and MMP-9 mRNA of human renal tubular epithelial cells HK-2 induced by TGF-β1, and explore its mechanism on prevention and treatment of renal fibrosis. Methods HK-2 cells were cultured by DMEM/F12 （1 ： 1） with 10% fetal bovine serum and divided into control group, TGF-[31 group （TGF-β1 10 ng/mL}, blank serum control group （TGF-β1 10 ng/mL＋ 10% blank serum）, Zhixiaotongmalning drug-containing serum intervention groups （TGF-β1 10 ng/mL＋ 10% low dose Zhixiaotongmaining, TGF-β1 10 ng/mL＋ 10% medium dose Zhixiaotongmalning, TGF-β1 10 ng/mL ＋ 10% high dose Zhixiaotongmalning）. After intervened for 24 h, the expression of CTGF, PAl-1 and MMP-9 mRNA were tested by fluorescence quantitatiye PCR assay. Results After the HK-2 cell was induced by TGF-β1, the expression of CTGF and PAl- 1 mRNA were increased and the expression of MMP-9 mRNA was decreased compared with the control （P〈0.05）. After intervened by Zhixiaotongrnaining, the expression of CTGF and PAl-1 mRNA decreased and the expression of MMP-9 mRNA increased compared with TGF-β1 group {P 〈0.05}. Conclusion Zhixiaotongmaining could prevent the development of renal fibrosis to some extent, its mechanism may be related with regulating the expression of CTGF, PAl- 1 and MMP-9 mRNA.