γ-分泌酶抑制剂干预对鼠心脏成纤维细胞平滑肌肌动蛋白-α及MMP-1/TIMP-1表达的影响  

The effects of DAPT intervention on the expression ofα-SMA and MMP-1/TIMP-1 in rats cardiac fibroblasts

作　　者：丁洁[1] 常煜胤 李文杰[2] 郭冰冰[2] 路明[2] 

机构地区：[1]徐州医学院研究生学院,江苏徐州221004 [2]徐州医学院附属医院儿科医院

出　　处：《临床心血管病杂志》2016年第7期735-738,共4页Journal of Clinical Cardiology

摘　　要：目的:探讨γ-分泌酶抑制剂(DAPT)抑制Notch信号通路对心肌成纤维细胞(CFs)平滑肌肌动蛋白-α(α-SMA)的表达及细胞外基质(ECM)降解平衡变化的影响。方法:采用胰酶消化法和差速贴壁法分离和纯化SD乳鼠CFs进行体外培养,将CFs分为正常组(N组,培养时不加干预药物)和尾加压素Ⅱ组(U组,加UⅡ10^(-8) mol/L)和DAPT+尾加压素Ⅱ组(D组,同时加入UⅡ10^(-8) mol/L和DAPT 75μmol/L),培养干预48h后采用RT-PCR法测基质金属蛋白酶-1(MMP-1)、组织金属蛋白酶抑制剂-1(TIMP-1)、α-SMA和Notch蛋白胞内段(Notch1)mRNA的表达;Western Blot检测α-SMA及Notch1蛋白的表达。结果:U组TIMP-1、α-SMA及Notch1的mRNA表达水平明显高于N组(均P<0.01),MMP-1mRNA的表达和MMP-1mRNA/TIMP-1mRNA比值低于N组(均P<0.01);D组TIMP-1、α-SMA、Notch1的mRNA表达以及α-SMA和Notch1蛋白的表达均较U组下调(均P<0.01),MMP-1mRNA的表达和MMP-1mRNA/TIMP-1mRNA比值较U组上调(P<0.01);但D组上述指标与N组相比差异无统计学意义。结论:DAPT可通过阻断Notch信号通路抑制尾加压素Ⅱ诱导的CFs表型转换,激活胶原降解途径,并可重调MMP-1/TIMP-1平衡,此作用有助于抑制心肌纤维化。Objective：To investigate the effect of DAPT blocking Notch signal pathway on the expression ofα-SMA and the degradation of extracellular matrix（ECM）on cardiac fibroblasts（CFs）.Method：The CFs of neonatal Sprague-Dawley rats were isolated with the method of trypsin digestion and differential anchoring velocity,then cultured in vitro.The generation 2-4of CFs were used for the experiment and randomly divided into control group（group N）：without drug intervention;urotensinⅡ（group U）：dealed with UⅡ10-8 mol/L;group DAPT＋ urotensinⅡ（group D）：with UⅡ10-8 mol/L and DAPT 75μmol/L.CFs were cultured for 48 h,then the expression of mRNA of matrix metalloproteinase-1（MMP-1）,tissue inhibitor of metalloproteinase-1（TIMP-1）,smooth muscle-Alpha excited protein（α-SMA）and the intracellular domain of Notch proteins（Notch1）were measured by RT-PCR,andα-SMA,Notch1 protein by Western Blot.Result：The expression of TIMP-1mRNA,α-SMA mRNA,Notch1 mRNA,α-SMA protein,and Notch1 protein were higher significantly（P0.01）,MMP-1mRNA expression and MMP-1mRNA/TIMP-1mRNA ratio were lower significantly（P0.01）in group U than in group N.Compared to those in group U,the expression of TIMP-1mRNA,α-SMA mRNA,Notch1 mRNA,α-SMA protein,and Notch1 protein were down-regulated significantly（P0.01）and MMP-1mRNA/TIMP-1mRNA ratio were up-regulated significantly（P0.01）in group D.But there were no statistically significant differences between group N and group D.Conclusion：DAPT can inhibit UⅡinduced CFs phenotype conversion by blocking the Notch signaling pathway,activate collagen degradation pathway,and regulate MMP-1/TIMP-1equilibrium.This effect is helpful to inhibit myocardial fibrosis.

关 键 词：心肌成纤维细胞 NOTCH信号通路 DAPT 尾加压素Ⅱ 

分 类 号：R542.2[医药卫生—心血管疾病]