MT1-MMP和PCSK9的联合抑制上调LDLR表达的研究  

Study on combined inhibition of MT1-MMP and PCSK9 up-regulating LDLR expression

作　　者：Adekunle Alabi 顾红梅 夏晓丹 张子洋 王贵清[2] 章大卫 Adekunle Alabi;GU Hongmei;XIA Xiaodan;ZHANG Ziyang;WANG Guiqing;ZHANG Dawei(Dept.of Pediatrics and Group on the Molecular and Cell Biology of Lipids,Faculty of Medicine and Dentistry,College of Health Sciences,University of Alberta,Edmonton,Alberta,Canada,T6G 2R3;Dept.of Orthopedics,Qingyuan People′s Hospital of the Sixth Affiliated Hospital of Guangzhou Medical University,Qingyuan 511518,China)

机构地区：[1]加拿大阿尔伯塔大学健康科学学院医学和牙科学院儿科和脂质分子和细胞生物学组,阿尔伯塔埃德蒙顿T6G 2R3 [2]广州医科大学附属第六医院清远市人民医院骨科,广东清远511518

出　　处：《华夏医学》2023年第1期18-25,共8页Acta Medicinae Sinica

基　　金：加拿大国立卫生研究院项目(PS155994,PS178091);加拿大自然科学和工程研究委员会项目(RGPIN-2016-06479)。

摘　　要：目的:探究Ⅰ型膜基质金属蛋白酶(MT1-MMP)和γ-分泌酶或前蛋白转化酶枯草溶菌素9(PCSK9)对低密度脂蛋白受体(LDLR)表达的影响。方法:应用DsiRNA分别沉默人肝细胞中MT1-MMP和PCSK9的表达并对细胞进行培养,选择DAPT抑制细胞中的γ-分泌酶,实时荧光定量PCR和免疫印迹法分别检测mRNA和蛋白质的表达水平。结果:MT1-MMP与γ-分泌酶或PCSK9的联合抑制可较单独抑制组进一步提高原代肝细胞中LDLR的表达水平;PCSK9^(-/-)小鼠再感染靶向小鼠MT1-MMP的shRNA腺相关病毒(AAV),会增加肝脏LDLR水平,降低血浆中的胆固醇浓度。结论:MT1-MMP和γ-分泌酶或PCSK9协同调节LDLR的表达。联合抑制MT1-MMP和γ-分泌酶或PCSK9可提高LDLR表达,并降低血浆中LDL胆固醇的水平,从而降低因现有手段无法有效控制血浆胆固醇水平的患者发生心血管病的风险。Objective:To investigate the effects of membrane type-1 matrix metalloproteinase(MT1-MMP)andγ-secretase or proprotein convertase subtilisin/kexin type 9(PCSK9)on the expression of low-density lipoprotein receptor(LDLR).Methods:DsiRNA was used to silence MT1-MMP and PCSK9 expression in cultured human hepatocytes,and the cells were cultured.DAPT was selected to inhibitγ-secretase in cells,and real-time fluorescence quantitative PCR and immunoblotting were used to measure mRNA and protein expression levels,respectively.Results:The combined inhibition of MT1-MMP andγ-secretase or PCSK9 further increased the expression level of LDLR in primary hepatocytes compared with the single inhibition group.The reinfection of PCSK9^(-/-)mice with shRNA adeno-associated virus(AAV)targeting MT1-MMP in mice increased the level of LDLR in liver and reduced the concentration of cholesterol in plasma.Conclusion:MT1-MMP andγ-secretase or PCSK9 synergistically regulate LDLR expression.Combined inhibition of MT1-MMP andγ-secretase or PCSK9 has the potential to further increase the expression of LDLR and reduce the level of plasma LDL cholesterol,thereby reducing the risk of cardiovascular disease in patients whose plasma cholesterol levels can not be successfully controlled by currently available strategies.

关 键 词：Ⅰ型膜基质金属蛋白酶 前蛋白转化酶枯草溶菌素9 低密度脂蛋白受体 联合抑制 

分 类 号：R54[医药卫生—心血管疾病]