乳鼠心肌细胞中C反应蛋白对基质金属蛋白酶-10表达调控的研究  

Effect of C-Reactive Protein on Matrix Metalloproteinase-10 Expression With the Underlying Mechanism in Cardiomyocyte of Neonatal Rats

作　　者：崔传珏[1] 魏英杰[1] 张秀芳[1] 史强[1] 胡盛寿[1] 

机构地区：[1]中国医学科学院北京协和医学院心血管病研究所阜外心血管病医院卫生部心血管疾病再生医学重点实验室,北京市100037

出　　处：《中国循环杂志》2010年第6期476-479,共4页Chinese Circulation Journal

基　　金：科技部"973"项目课题(2010CB529506)

摘　　要：目的:研究在乳鼠心肌细胞中C反应蛋白诱导基质金属蛋白酶-10(MMP-10)表达的作用及其分子机制。方法:培养乳鼠心肌细胞,以炎性因子C反应蛋白诱导MMP-10的表达。分别应用酶谱法和实时定量反转录—聚合酶链反应检测培养上清中MMP-10的活性和细胞中MMP-10信使核糖核酸(mRNA)表达水平的变化,应用蛋白免疫印迹杂交技术观察胞外调节激酶(ERK)通路以及转录因子活化蛋白-1的蛋白水平的变化。实验分为4组:对照组,C反应蛋白组,C反应蛋白+抑制剂组和抑制剂组。结果:心肌细胞给予C反应蛋白(5μg/ml)刺激后24 h,细胞培养上清中MMP-10活性较对照组升高了9.23倍(P<0.01);细胞MMP-10 mRNA水平与0 h相比,6 h增加了0.83倍(P<0.05),12 h达最高,增加了2.67倍(P<0.05),24 h略有下降,增加了1.92倍(P<0.05),差异均有统计学意义。C反应蛋白可激活心肌细胞p-ERK1和p-ERK2,从15 min开始,可持续到240 min;C反应蛋白刺激细胞4 h,活化蛋白-1的蛋白水平与0 h相比,增加了3.66倍(P<0.01),差异有统计学意义;ERK抑制剂PD98059预处理心肌细胞,可阻断C反应蛋白引起的活化蛋白-1的蛋白水平的上调作用以及MMP-10活性的增加及MMP-10 mRNA的表达。结论:C反应蛋白通过ERK通路,上调转录因子活化蛋白-1的蛋白水平,从而对心肌细胞中MMP-10基因的转录和表达进行调节。Objective： To investigate the effect of C-reactive protein（CRP） on matrix metalloproteinase-10（MMP-10） expression with the underlying mechanism.Methods：The experiments were divided into 4 groups,Control group： the neonatal rat cardiomyocytes were regularly cultured;CRP group： 5μg /ml of CRP was added into the cell culture medium;CRP plus inhibitor group： CRP and 50μM of PD98059 were added into cell the culture medium;and Inhibitor group： PD98059 was added.The mRNA expression of MMP-10 was analyzed by real-time RT-PCR.The protein levels of MMP-10,extracellular regulated protein kinases（ERK） and activated protein-1 were measured with Western blot and zymographic assay respectively.Results：By 24 hours of treatment,compared with Control group,the activity of MMP-10 increased 9.23 times in CRP group（P0.01） in the conditioned medium;while by 6h,12h,and 24h of culture,the mRNA level of MMP-10 increased 0.83 times,2.67 times,and 1.92 times in CRP group（P0.05 respectively）.CRP could activate the ERK1 and ERK2 signaling pathway started from 15 min of culture,and this effect lasted for 240 min.By 4h of treatment,the protein level of activated protein-1 increased 3.66 times in CRP group（P0.01）.The ERK inhibitor,PD98059,could block the increased mRNA expression and enzyme activity of MMP-10,inhibit the increased protein level of activated protein-1 which were induced by CRP.Conclusion： CRP could regulate the mRNA expression of MMP-10,that was mediated by ERK signaling pathway and activated protein-1.

关 键 词：C反应蛋白 心肌细胞 基质金属蛋白酶-10 胞外调节激酶通路 活化蛋白-1 

分 类 号：R541[医药卫生—心血管疾病]