氧化低密度脂蛋白增加巨噬细胞基质金属蛋白酶-9和-12的表达  

Oxidized low-sensity lipoprotein increases expression of matrix metalloproteinase-9 and-12

作　　者：何春燕[1] 周新[2] 武军驻[1] 戴赵明[1] 王贞[1] 洪嘉玲[1] 

机构地区：[1]武汉大学医学院生化与分子生物学系,武汉430071 [2]武汉大学中南医院检验科,武汉430071

出　　处：《基础医学与临床》2003年第2期160-164,共5页Basic and Clinical Medicine

基　　金：湖北省科技厅攻关课题资助项目 (2 0 0 0 - 4 - 2 7)

摘　　要：为探讨巨噬细胞在动脉粥样斑块不稳定性中的作用机制 ,观察致动脉粥样硬化的主要炎性因子 氧化低密度脂蛋白 (oxLDL) ,对巨噬细胞基质金属蛋白酶MMP 9和MMP 12表达和分泌的影响。在体外分离培养小鼠腹腔巨噬细胞 ,以不同浓度低密度脂蛋白 (LDL)、氧化低密度脂蛋白 (oxLDL)作用细胞 ;作用 6h后用RT PCR检测MMP 9,MMP 12mRNA表达 ,作用 2 4h后用Westernblot和 β 酪蛋白酶谱法检测MMP 9,MMP 12酶蛋白的分泌及活性。ox LDL增强巨噬细胞MMP 9,MMP 12mRNA表达和酶蛋白的分泌及活性 ,对MMP 12的诱导作用更为显著且呈浓度依赖性。表明oxLDL可能通过同时上调巨噬细胞基质金属蛋白酶MMP 9和MMPMatrix Metalloproteinase 9 and Matrix Metalloproteinase 12(MMP 9,MMP 12) produced by macrophages have been shown to play critical roles in atherosclerosis through impacts on vascular remodeling and plaque disruption . The present study was undertaken to investigate the effect of oxidized low density lipoprotein(ox LDL) which was implicated in many proatherogenic events, on the expression of MMP 9 and MMP 12 in an in vitro murine macrophage system . The macrophages were incubated with various concentrations of LDL, copper oxidized LDL. After incubation for 6 hrs, RT PCR analysis was used to measure the mRNA exression of MMP 9 and MMP 12 . After incubation for 24 hrs, media from the plates were immediately taken and stored at 80℃ for the checking of enzymatic activity of MMP 12 by β casein zymography and for determination of protein levels of MMP 9 by Western blot . Exposure to ox LDL (10 to 50 mg/L) increased MMP 9 mRNA and protein expression . OxLDL stimulated macrophages produced increased casein degrading activities at both 22 and 29 Kda, and ox LDL also significantly increased the mRNA level of MMP 12 in a dose dependent manner with a peak (6.64 fold versus control) at 50 mg/L ox LDL . LDL(80mg/L) had no significant effect on MMP 9 and MMP 12 expression . These data demonstrated that ox LDL contributed to macrophage mediated matrix breakdown in the atherosclerotic plaques by upregulating MMP 9 and MMP 12 expression by macrophages.

关 键 词：动脉粥样硬化 不稳定性斑块 氧化低密度脂蛋白 巨噬细胞基质金属蛋白酶MMP-9 

分 类 号：R363[医药卫生—病理学]