机械牵张力改变诱导YAP蛋白下调参与升主动脉瘤的发生  

Down regulation of YAP protein induced by the change of mechanical stress involves in the pathogenesis of ascending aortic aneurysm

作　　者：姜文剑[1] 李海洋[1] 王晓龙[1] 张宏家[1] JIANG Wenjian;LI Haiyang;WANG Xiaolong;ZHANG Hongjia(Department of Cardiac Surgery,Beijing Anzhen Hospital,Capital Medical University,Beijing Institute of Heart,Lung and Blood Vessel Diseases,Beijing 100029,China)

机构地区：[1]首都医科大学附属北京安贞医院-北京市心肺血管疾病研究所心脏外科北京市心肺血管疾病研究所心血管疾病精准医学北京实验室北京大血管疾病诊疗中心北京市大血管外科植入式人工材料工程技术研究中心,100029

出　　处：《心肺血管病杂志》2019年第1期85-92,共8页Journal of Cardiovascular and Pulmonary Diseases

基　　金：国家自然科学基金(81800404;81470580);心血管疾病精准医学北京实验室(PXM2017_014226_000037);北京市优秀人才培养资助青年骨干个人项目(2017000021469G254);首都卫生发展科研专项项目-青年优才(2018-4-2068);北京市医院管理局青年人才培养"青苗"计划(QML20180601)

摘　　要：目的:研究YAP蛋白在升主动脉瘤(AAA)的发病中是否发挥作用。方法:研究对象由2部分组成:①AAA患者:收集2014年9月至2015年9月,于首都医科大学附属北京安贞医院接受单纯升主动脉替换或者合并升主动脉替换手术的AAA患者以及接受心脏移植手术患者的标本;②使用cyclic stretch拉伸仪拉伸细胞以构建细胞凋亡模型。应用弹力板染色、免疫组化、免疫荧光和Tunel染色观察主动脉壁组织的形态学,Western blotting检测主动脉壁组织及主动脉平滑肌细胞株中YAP在蛋白水平的表达。结果:①弹力板染色、免疫组化和Tunel染色结果显示:AAA患者主动脉壁组织细胞外基质异常,且主动脉壁血管平滑肌细胞(VSMCs)凋亡增多。②免疫组化染色和免疫荧光染色结果显示:AAA患者主动脉壁组织中VSMCs凋亡增多,免疫荧光染色的统计分析结果显示:AAA患者的主动脉壁组织中VSMCs的凋亡较正常主动脉壁组织显著增多(P<0.05)。③Western blotting检测、免疫组化染色和免疫组化染色结果显示,与正常主动脉壁组织相比,AAA患者主动脉壁组织中血管平滑肌中的YAP蛋白表达量显著降低;计算机软件分析Western blotting结果的蛋白灰度显示YAP蛋白的表达量降低近90%,统计结果显示差异有统计学意义(P<0.05);免疫荧光染色结果显示:AAA患者的主动脉壁组织中VSMCs的凋亡较正常主动脉壁组织显著增多(P<0.05)。④Western blotting检测结果显示:相对于正常对照组,CRL-1999人主动脉平滑肌细胞株接受Cyclic Stretch机械牵拉36h后,细胞中的YAP蛋白的表达量显著减少,提示人主动脉平滑肌细胞受到机械牵张后会使细胞内的YAP表达量降低,进而发生凋亡。结论:YAP在AAA的发病中发挥重要作用。当升主动脉细胞外基质发生变化时,机械牵张力的改变可以使YAP的表达下调,进而通过促进主动脉血管平滑肌的凋亡参与AAA的发病过程。Objective:The purpose of this study was to investigate the expression of YAP in aortic wall tissue of human ascending aortic aneurysm,and to investigate the mechanism of ascending aortic aneurysm.Methods:The materials of this study were consisted by 2 parts:1Patients with ascending aortic aneurysm or cardiac transplantation.2The cell apoptosis model was constructed by using stretch cyclic stretching apparatus.The applications of elastic plate staining,immunohistochemistry,immunofluorescence and Tunel staining were used to detect the morphology of aortic wall tissue.And the Western blotting was used to detect the expression of YAP in the human aorticwall and the cell line of human aortic smooth muscle cells.Results:1The result of elastic plate staining,immunohistochemistry and Tunel staining:extracellular matrix of aortic wall tissue was abnormal in patients with ascending aortic aneurysm,and the apoptosis of vascular smooth muscle cells was increased.2The result of immunohistochemical staining and immunofluorescence staining:The apoptosis of vascular smooth muscle cells was significantly increased in patients with ascending aortic aneurysm(P<0.05).3The result of Western blotting,immunohistochemical staining and immunohistochemical staining:The expression of YAP was significantly decreased in aortic wall tissue of patients with ascending aortic wall tissue(P<0.05).4The result of Western blotting:Compared with the control group,the expression of YAP in cells after 36-hour cyclic stretch was significantly decreased,which indicated that the expression of YAP in human aortic smooth muscle cells will reduce after cyclic stretch,and then promote the apoptosis of aortic smooth muscle cells.Conclusions:YAP plays an important role in the pathogenesis of ascending aortic aneurysm.When the extracellular matrix of the ascending aorta was abnormal,the change of mechanical stress will make the expression of YAP down regulated,and promote the apoptosis of the aortic vascular smooth muscle,both of which were involved in the patho

关 键 词：YAP蛋白 升主动脉瘤 细胞外基质 血管平滑肌细胞 凋 

分 类 号：R54[医药卫生—心血管疾病]