摘要
目的:探讨睾丸特异性Y编码蛋白5(TSPYL5)在心肌缺血损伤中的作用及机制。方法:在体内实验中,构建心肌缺血小鼠模型,采用实时荧光定量聚合酶链反应(qRT-PCR)、Western blot和免疫组织化学染色等方法检测正常对照组和心肌缺血组小鼠心肌组织中TSPYL5的表达情况。在体外实验中,将HL-1细胞分成4组:空白对照组、TSPYL5过表达组、化学缺氧组和TSPYL5过表达+化学缺氧组,采用流式细胞仪检测凋亡率,Western blot检测活化胱天蛋白酶-3(cleaved caspase-3)、Bcl-2和p53的蛋白表达水平,qRT-PCR检测Bax和Bcl-2的mRNA表达水平。结果:与正常对照组相比,心肌缺血组小鼠心肌TSPYL 5的mRNA和蛋白表达水平均显著下降(P均<0.05)。使用500μmol/L氯化钴能够成功诱导HL-1化学缺氧,与空白对照组相比,化学缺氧组凋亡细胞比例和cleaved caspase-3、p53的蛋白表达水平显著升高,Bcl-2的mRNA和蛋白表达水平显著下降,Bax的mRNA表达水平显著升高;与化学缺氧组相比,TSPYL5过表达+化学缺氧组cleaved caspase-3、p53蛋白水平显著下降,Bcl-2的mRNA和蛋白表达水平显著升高,Bax的mRNA表达水平显著下降(P均<0.05)。结论:TSPYL5能够通过抑制p53通路减轻缺氧诱导的心肌细胞凋亡。
Objective:To investigate the role and mechanism of testis-specific Y-encoded-like protein 5(TSPYL5)in myocardial ischemia injury.Methods:The myocardial ischemia model of mice was established.The expression level of TSPYL5 in myocardial tissue of mice in control group and myocardial ischemia group was detected by qRT-PCR,Western blot and immunohistochemistry.In vitro study,HL-1 cells were divided into four groups:blank control group,TSPYL5 overexpression group,chemical hypoxia group,and TSPYL5 overexpression+chemical hypoxia group.The apoptotic rate was detected by flow cytometry,and the protein expression levels of cleaved caspase-3,Bcl-2 and p53 were detected by western blot.The mRNA expression levels of Bax and Bcl-2 were detected by qRT-PCR.Results:Compared with control group,TSPYL5 mRNA and protein expression levels in myocardial ischemia group were significantly decreased(all P<0.05).500μmol/L CoCl 2 could successfully induce HL-1 chemical hypoxia.Compared with the blank control group,the proportion of apoptotic cells in the chemical hypoxia group was significantly increased,the protein expression levels of cleaved caspase-3 and p53 were significantly increased,and the mRNA and protein expression levels of Bcl-2 were decreased,and the mRNA expression level of Bax was significantly increased(all P<0.05).Compared with chemical hypoxia group,the levels of cleaved caspase-3 and p53 protein in TSPYL5 overexpression+chemical hypoxia group were significantly decreased,the mRNA and protein expression levels of Bcl-2 were significantly increased,and the level of Bax mRNA was significantly decreased(all P<0.05).Conclusions:TSPYL5 can alleviate hypoxia induced apoptosis in cardiomyocytes by inhibiting p53 signaling pathway.
作者
吴佳伟
范怡伦
丁丹
薛松
黄日太
WU Jiawei;FAN Yilun;DING Dan;XUE Song;HUANG Ritai(Department of Cardiovascular Surgery,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200120,China)
出处
《国际心血管病杂志》
2021年第4期236-240,共5页
International Journal of Cardiovascular Disease
关键词
心肌缺血
睾丸特异性Y编码蛋白5
缺氧
P53
Myocardial ischemia
Testis-specific Y-encoded-like protein 5
Hypoxia
p53
