姜黄素抑制HSC—T6细胞迁徙和侵袭的机制

Curcumine inhibits migration and invasion of hepatic stellate cells by reducing MMP-2 expression and activity

目的探讨姜黄素抑制肝星状细胞株HSC—T6迁徙和侵袭的分子机制。方法培养大鼠肝星状细胞株HSC-T6，并以刀豆蛋白A（ConA）激活，以不同剂量姜黄素处理后，用Westem blot检测HSC—T6基质金属蛋白酶2（MMP-2）的表达水平；明胶酶谱法检测其活性。同时检测HSCT6活化前后的迁徙和侵袭情况。数据采用单因素方差分析。结果静息状态下HSC—T6仅表达少量MMP2，ConA激活后MMP-2表达水平显著增加；25、50、100μmol／L姜黄素处理后，MMP-2表达水平分别减少了21．8％±5．1％、65．5％±9．2％和87．9％±11．5％，P值均〈0．05。明胶酶谱实验显示静息状态下HSC—T6表达少量MMP-2且几乎没有活性，ConA激活后MMP-2表达水平和活性显著升高；不同剂量姜黄素处理后，其表达水平和活性也呈剂量依赖性降低，P值均〈0．01。静息状态下HSC-T6很少发生迁徙和侵袭，ConA激活后HSC—T6的迁徙力和侵袭力显著增强；25、50、100μmol／L姜黄素处理后，HSC-T6迁徙数分别减少了27．5％±5．8％、54．4％±7．6％和67．1％±9．3％（P值均〈0．01），HSC—T6侵袭细胞数也呈剂量依赖性减少（P值均〈0．05）。结论姜黄素可能通过下调MMP-2的表达水平及其活性来抑制HSC—T6的迁徙和侵袭，进而抑制肝纤维化。

Objective To investigate the molecular mechanism of the inhibitory effect of curcumine on the migration and invasion of hepatic stellate cells （HSC）. Methods Rat hepatic stellate cells were cultured and activated with ConA. Matrix metalloproteinase-2 （MMP-2） expression and activity was determined by Western blot and gelatin zymography. Migration and invasion of HSC was assessed by wound healing assay and modified Boyden chamber assay. Results Curcumine reduced the level and activity of MMP-2 expression in activated HSC in a dose-dependent manner. When treated with 25, 50 or 100 μ mol/L curcumine, the expression of MMP-2 was reduced by 21.8% ±5.1%, 65.5% ±9.2% or 87.9% ±11.5% （P 〈 0.05）, and the activity of MMP-2 was also significently reduced by curcumine. Migration and invasion of activated HSC was also inhibited by curcumine in a dose-dependent way. When treated with 25, 50 or 100 μmol/L curcumine, the migration of activated HSC was reduced by 27.5% ±5.8%, 54.4% ±7.6% or 67.1% ±9.3% （P 〈 0.05）, and the invasion of activated HSC was also significently reduced by curcumine. Conclusion Curcumine inhibits migration and invasion of activated HSC by reducing MMP-2 expression and activity.