丝氨酸蛋白酶Omi/HtrA2表达增加促进心肌衰老的作用

doi:10.3969/j.issn.1006-7795.2021.05.014

摘要/Abstract

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。

关键词: Omi/HtrA2, 心肌, 衰老

Abstract: 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.

Key words: Omi/HtrA2, myocardium, aging

中图分类号:

R542.2

隗歆, 刘丹, 武烨, 吕坤译, 申言, 刘慧荣. 丝氨酸蛋白酶Omi/HtrA2表达增加促进心肌衰老的作用[J]. 首都医科大学学报, 2021, 42(5): 783-789.

Wei Xin, Liu Dan, Wu Ye, Lyu Kunyi, Shen Yan, Liu Huirong. Omi/HtrA2 overexpression is associated with myocardial aging[J]. Journal of Capital Medical University, 2021, 42(5): 783-789.

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