丝氨酸蛋白酶Omi/HtrA2表达增加促进心肌衰老的作用  

Omi/HtrA2 overexpression is associated with myocardial aging

作　　者：隗歆 刘丹 武烨[1,2] 吕坤译 申言 刘慧荣 Wei Xin;Liu Dan;Wu Ye;Lyu Kunyi;Shen Yan;Liu Huirong(Department of Physiology and Pathophysiology,School of Basic Medical Sciences,Capital Medical University,Beijing 100069,China;Beijing Key Laboratory of Metabolic Disturbance Related Cardiovascular Disease,Beijing 100069,China;Yan Jing Medical College,Capital Medical University,Beijing 101300,China)

机构地区：[1]首都医科大学基础医学院生理学与病理生理学系,北京100069 [2]代谢紊乱相关心血管疾病北京市重点实验室,北京100069 [3]首都医科大学燕京医学院,北京101300

出　　处：《首都医科大学学报》2021年第5期783-789,共7页Journal of Capital Medical University

基　　金：国家自然科学基金(81700270);北京市自然科学基金(7192019);首都医科大学燕京医学院学院基金(19qdky02)。

摘　　要：目的观察线粒体促凋亡蛋白Omi/HtrA2表达增加对心肌衰老的影响。方法构建心肌特异性过表达Omi/HtrA2小鼠模型,通过Western blotting法检测衰老标志物p53、p21、p16表达水平;分析Omi/HtrA2与衰老标志物表达的相关性;构建Omi/HtrA2稳转H9c2细胞系,通过Western blotting法检测Omi/HtrA2表达增加后衰老指标p53、p21、p16的改变;通过β-半乳糖苷酶(β-gal)染色比较β-半乳糖苷酶染色阳性衰老细胞的改变;通过Omi/HtrA2特异性抑制剂UCF101抑制Omi/HtrA2功能,检测抑制Omi/HtrA2活性后衰老标志物的表达水平及阳性衰老细胞变化。结果(1)与对照组相比,心肌特异性过表达Omi/HtrA2小鼠心肌组织中,Omi/HtrA2表达量显著增加(P<0.01);过表达Omi/HtrA2心肌组织衰老标志物p53、p21、p16增加(P<0.01);(2)在心肌组织中Omi/HtrA2表达量与衰老标志物p53、p21、p16呈正相关(P<0.05);(3)与H9c2细胞(control组)相比,Omi/HtrA2稳转H9c2细胞(Omi组)衰老标志物p53、p21、p16表达增加(P<0.05);β-半乳糖苷酶染色结果显示,Omi组衰老细胞增加;与Omi组相比,Omi/HtrA2特异性抑制剂UCF101处理组(Omi+UCF101组)衰老标志物p53、p21、p16表达降低(P<0.05);β-半乳糖苷酶染色阳性细胞减少。结论Omi/HtrA2的表达与衰老标志物呈正相关,表达增加的Omi/HtrA2促进心肌衰老。Objective To observe the effect of increased serine protease Omi/HtrA2 on myocardial aging.Methods Overexpressed cardiac-specific Omi/HtrA2 transgenic mice was constructed,the expression of senescence markers p53,p21 and p16 were detected by Western blotting,and the correlation between Omi/HtrA2 and the senescence markers was analyzed.The aging indicators were detected in overexpressed Omi/HtrA2 H9c2 cells(Omi group)by Western blotting.The senescent cells were detected by β-galactosidase staining.Compared with overexpressed Omi/HtrA2 cells,the expression of senescence markers and the β-galactosidase positive senescent cells were detected after inhibiting the function of Omi/HtrA2 by Omi/HtrA2 specific inhibitor UCF101.Results(1)Compared with the control group,the expression of Omi/HtrA2 was significantly increased in the heart of myocardium-specific overexpressed Omi/HtrA2 transgenic mice(P<0.01).The aging markers p53,p21 and p16 were significantly increased in Omi/HtrA2 overexpressed group(P<0.01).(2)The expression of Omi/HtrA2 was positively correlated with the aging markers p53,p21,and p16(P<0.01).(3)Compared with H9c2 cells(control group),expression of the senescence markers p53,p21 and p16 and the β-galactosidase positive senescent cells increased in Omi group(P<0.05).Compared with Omi group,expression of the senescence markers and the β-galactosidase positive senescent cells decreased in Omi/HtrA2 H9c2 cells treated by Omi/HtrA2 specific inhibitor UCF101(Omi+UCF101 group)(P<0.05).Conclusion The expression of Omi/HtrA2 is positively correlated with aging markers.Overexpression of Omi/HtrA2 promoted myocardial aging.

关 键 词：OMI/HTRA2 心肌 衰老 

分 类 号：R542.2[医药卫生—心血管疾病]