缺氧肝细胞对肝星状细胞基质金属蛋白酶-2表达及活性水平的影响  

Effects of hypoxic hepatocytes on the expression and the activity of matrix metalloproteinase-2 in rat hepatic stellate cells

作　　者：李静[1] 陈平圣[1] 

机构地区：[1]东南大学医学院病理学与病理生理学系,江苏南京210009

出　　处：《东南大学学报（医学版）》2010年第3期272-275,共4页Journal of Southeast University(Medical Science Edition)

基　　金：国家自然科学基金资助项目(30470780)

摘　　要：目的:探讨缺氧肝细胞对大鼠肝星状细胞基质金属蛋白酶-2(MMP-2)表达及其活性的影响。方法:缺氧培养肝细胞BRL-3A(氧浓度5%)12 h后取其培养上清作为条件培养基,培养肝星状细胞HSC-T6,设6、12、24 h 3个时间点,RT-PCR、酶图法分别检测HSC-T6 MMP-2 mRNA表达及其酶活性。以无血清DMEM培养肝星状细胞作为对照组。结果:随条件培养时间的延长,缺氧条件培养组HSC-T6 MMP-2活性均升高(30.74±8.22、31.71±8.10、49.79±13.61),且高于对照组(18.58±1.62、25.98±2.73、32.14±3.76),但扣除本底后缺氧条件培养基组HSC-T6 MMP-2活性(12.99±2.05、14.01±2.00、29.44±3.62)均低于相应对照组,在6 h和12 h时间点差异有统计学意义(P6 h=0.039,P12 h=0.007),缺氧条件培养基组HSC-T6 MMP-2 mRNA表达量(0.44±0.07、3.66±1.10、4.29±0.99)高于相应对照组(0.61±0.07、2.50±0.27、0.52±0.16),差异具有统计学意义(P6 h=0.044,P24 h=0.030)。结论:物理缺氧条件下,肝细胞分泌或释放了可溶性物质作用于肝星状细胞,使其MMP-2表达上调,活性下降。肝细胞分泌或释放了何种物质尚待进一步研究。Objective：To investigate the effects of hypoxic hepatocytes on the expression and the activity of matrix metalloproteinase-2（MMP-2） in rat hepatic stellate cells.Methods： Hepatocytes（BRL-3A） were cultured in Dulbecco′s modified eagle medium with 10% fetal calf serum at 37 ℃ and 5% CO2.When reaching confluence,the hepatocytes were incubated in serum-free medium and put into the hypoxia-box,in which the concentration of oxygen was 5%.After 12 h,the supernatants were harvested,and were used to culture rat hepatic stellate cells（HSC cell line） for 6 h,12 h,24 h respectively.And then the supernatants and the cells were harvested.The expression of the MMP-2 mRNA in HSC cells was detected by RT-PCR.The activity of the MMP-2 in the supernatant was detected by zymography.Results：The expression of MMP-2 in the hypoxic conditional groups at three time points（0.44±0.07,3.66±1.10,4.29±0.99） were higher than those in the control groups（0.61±0.07,2.50±0.27,0.52±0.16）,P6 h=0.044,P24 h=0.030.However the activity of MMP-2 in the hypoxic conditional groups（12.99±2.05,14.01±2.00,29.44±3.62） were lower than those in the control groups（18.58±1.62,25.98±2.73,32.14±3.76）,P6 h=0.039,P12 h=0.007.Conclusion：Hypoxic hepatocytes may secret some substances to promote MMP-2 expression but depress the activity in hepatic stellate cells,and the effect correlated with the time.It is worth to studying further that what the substances are.

关 键 词：基质金属蛋白酶-2 缺氧 肝细胞 肝星状细胞 相互作用 

分 类 号：R-33[医药卫生]