去泛素化酶通过肿瘤坏死因子-α途径对小鼠心肌衰老的作用  

Cylindromatosis protects against myocardial senescence in mice via the tumor necrosis factor-αpathway

作　　者：滕敬华[1] 胡恋 程飞[1] 何茜[1] 王洁[1] 丁妍 王治校[1] Teng Jinghua;Hu Lian;Cheng Fei;He Qian;Wang Jie;Ding Yan;Wang Zhixiao(Department of Cardiology,Taihe Hospital,Hubei University of Medicine,Shiyan 442000,China)

机构地区：[1]十堰市太和医院、湖北医药学院附属太和医院心血管内科,十堰442000

出　　处：《中华生物医学工程杂志》2023年第2期129-134,共6页Chinese Journal of Biomedical Engineering

基　　金：国家自然科学基金面上项目(82270530);湖北省自然科学基金创新群体项目(2022CFA036);湖北医药学院研究生创新训练项目(YC2019013,YC2019018)。

摘　　要：目的探讨去泛素化酶(CYLD)在小鼠心肌细胞衰老中的作用及机制。方法选取野生型FVB(WT)小鼠和CYLD过表达(CYLD+/+)小鼠构建自然衰老模型。2月龄雄性WT小鼠(WT-2M,n=10)和CYLD+/+小鼠(CYLD-2M,n=10)作为年轻组,普通饲养17.5月(WT-17.5M,CYLD-17.5M,n=10)作为年老组。小动物超声检测心功能[左室短轴缩短率(LVFS)、左室射血分数(LVEF)],蛋白免疫印迹检测心肌组织衰老相关蛋白p53、p21、p16表达水平,实时荧光定量PCR(qPCR)检测心肌组织衰老相关分泌表型(SASP)[细胞因子白介素-1β(IL-1β)、基质金属蛋白酶3(MMP3)、趋化因子CXCL1(CXCL1)]转录水平。转录组测序检测CYLD调控心肌细胞衰老的信号通路,对肿瘤坏死因子α(TNF-α)通路行qPCR验证。结果与WT-17.5M小鼠比较,CYLD-17.5M小鼠心功能LVEF和LVFS值增高(均P<0.05);p53、p21、p16蛋白表达水平减低(均P<0.05),IL1B、MMP3、CXCL1转录水平也下降(均P<0.05)。与WT-2M小鼠比较,WT-17.5M小鼠p53、p21、p16蛋白表达明显升高(均P<0.01),IL1B、MMP3及CXCL1转录水平上调(均P<0.01)。转录组测序发现WT-17.5M小鼠较WT-2M小鼠表达上调的信号通路64个,CYLD-17.5M小鼠较WT-17.5M小鼠表达下调的信号通路37个,二者共有的差异信号通路29个。选取TNF-α通路行qPCR验证,WT-17.5M较WT-2M小鼠TNF-α转录水平明显上调(P<0.05),且高于CYLD-17.5M小鼠(P<0.05)。结论CYLD对心肌自然衰老起保护作用,TNF-α通路介导可能是CYLD保护心肌衰老的作用机制之一。Objective To investigate the role of the deubiquitinase Cylindromatosis(CYLD)in aging of mouse cardiomyocytes and the underlying mechanism.Methods Wild-type(WT)FVB mice and CYLD-overexpressing(CYLD+/+)mice were used to establish mice models of natural aging.Then,2-month-old male wild-type FVB mice(WT-2M,n=10)and CYLD+/+mice(CYLD-2M,n=10)were assigned to the young age group,and those routinely raised for 17.5 months(WT-17.5M,CYLD-17.5M,n=10 each)to the old age group.Small animal ultrasonography was used to measure the cardiac function(LVEF,LVFS),Western blotting was used to examine the expression of aging-related proteins p53,p21,and p16 in myocardial tissues,and qPCR was used to determine the transcription of senescence associated secretory phenotypes(SASP)interlenkin-1β(IL-1β),matrix metalloproteinase 3(MMP3)and CXC chemokine ligand 1(CXCL1)in myocardial tissues.Transcriptome sequencing was used to detect the signaling pathways by which CYLD regulates cardiomyocyte senescence.Tumor necrosis factor-α(TNF-α)pathway was verified by qPCR.Results Compared with WT-17.5M mice,CYLD-17.5M mice presented better cardiac function with higher values of LVEF and LVFS(all P<0.05),lower expression levels of p53,p21,and p16 proteins(all P<0.05),and less transcription of IL1B,MMP3,and CXCL1(all P<0.05).Compared with WT-2M mice,the WT-17.5M mice had significantly higher expression of p53,p21 and p16 proteins(all P<0.01),and up-regulated transcription levels of IL1B,MMP3 and CXCL1(all P<0.01).Transcriptome sequencing showed 64 up-regulated signaling pathways in WT-17.5M mice when comparing to WT-2M mice,and 37 down-regulated signaling pathways in CYLD-17.5M mice when comparing to WT-17.5M mice;there were 29 pathways with differential signaling when comparing CYLD-17.5M to WT-17.5M mice.qPCR with TNF-αpathway selected for verification showed significantly increased TNF-αtranscription in WT-17.5M mice than in WT-2M mice and CYLD-17.5M mice(all P<0.05).Conclusion CYLD plays a protective role in natural cardiomyocyte senescence.M

关 键 词：去泛素化酶 小鼠 心肌细胞 衰老 肿瘤坏死因子-Α 

分 类 号：R542.2[医药卫生—心血管疾病]