螺内酯对肝星状细胞表达基质金属蛋白酶及其抑制剂的影响

Effect of spironolactone on expression of matrix metalloproteinases and tissue inhibitor of matrix metalloproteinase-1 in hepatic stellate cells

目的研究螺内酯干预肝星状细胞(HSCs)后基质金属蛋白酶2(MMP-2)、MMP-9、MMP-13和其组织抑制剂1(TIMP-1)的变化。探讨螺内酯对胶原代谢影响的机制。方法应用不同浓度螺内酯(1×10-4)mol/L^(1×10-7)mol/L干预HSCs 48小时,采用反转录-聚合酶链反应(RT-PCR)方法检测MMP-2、MMP-9、MMP-13 mRNA和TIMP-1 mRNA的表达。结果①螺内酯组的MMP-13基因表达强度上调,螺内酯(1×10-4)mol/L^(1×10-6)mol/L浓度组MMP-13基因表达强度(0.91±0.13、0.80±0.01、0.67±0.15)均明显高于对照组(0.53±0.10)(P<0.01)。1×10-4mol/L浓度组MMP-13基因表达强度接近对照组的2倍。②螺内酯干预HSCs 48小时后,TIMP-1基因表达减少,螺内酯(1×10-4)mol/L^(1×10-6)mol/L浓度组(0.15±0.05、0.28±0.15、0.37±0.03)明显低于对照组(0.47±0.04)(P<0.01或<0.05)。③螺内酯不同浓度干预HSCs 48小时后,HSCs MMP-2、MMP-9基因表达无明显变化(P>0.05)。结论螺内酯可抑制HSCs TIMP-1基因的表达,增加间质胶原酶MMP-13 mRNA的含量。螺内酯可能通过对MMPs及TIMP-1影响而促进胶原降解。

Objective To investigate the effects of spironolaetone on expression of metalloproteinases （MMP-2, MMF-9,MMP-13 ） and tissue inhibitor of matrix metalloproteinase-1 （TIMP-1） in hepatic stellate cells（HSCs）. And to study the mechanism how spironolactone affects the collagen metabolism in HSCs. Metheds The activated HSCs were treated with different concentrations of spironolactone（1×10^-4- 1×10^-4mol/L） for 48 hours. The expressions of MMF-2,MMP-9,MMP-13 and TIMP-1 mRNA were evaluated by RT-PCR（MMPs or TIMP-1/β-actin）. Results The expression of MMP-13mRNA in spironolactone groups was up-regulated. This gene expression in 1 ×10^-4 - 1 ×10^-5 mol/L spironolactone treated groups （0.91 ± 0.13,0.80 ± 0.01 and 0.67 ± 0. 15 respectively） was significantly higher than that in the control,0.53±0.10（ P 〈0.01）. Treatment with spironolactone for 48 hours, the expression of TIMP-1 mRNA decreased dose-dependently,and among 1×10^-4 mol/L and 1 ×10^-6 mol/L groups,the decrease was the most obvious （0.15±0.05,0.28±0.15 and 0. 37±0.03 respectively vs control group 0. 47±0.04, P 〈0.01 or P 〈0.05）. MMP-2 mRNA and MMP-9 mRNA expression in HSCs stayed unchanged after incubation with different concentrations of spironolactone for 48 hours. Conclusion Spironolactone may inhibit the expression of TIMP-1 and increase MMP-13 mRNA. Spironolactone may promote the collagen degradation by regulating MMFs and TIMP-1 expression.