钙感受器STIM1参与主动脉平滑肌细胞衰老的作用研究  

Role of calcium sensor STIM1 in senescence of aortic smooth muscle cells

作　　者：曾鹏 秦晓玥 王义蓉 李穗敏 陈淑贞 邝素娟 杨慧[3,4] 饶芳 邓春玉[1,2,3,4] ZENG Peng;QIN Xiao-yue;WANG Yi-rong;LI Sui-min;CHEN Shu-zhen;KUANG Su-juan;YANG Hui;RAO Fang;DENG Chun-yu(School of Medicine,South China University of Technology,Guangzhou 510006,China;School of Biological Science and Engineering,South China University of Technology,Guangzhou 510006,China;Guangdong Provincial Key Laboratory of Clinical Pharmacology,Medical Research Center of Guangdong Provincial People's Hospital,Guangdong Academy of Medi-cal Sciences,Guangzhou 510080,China;Cardiovascular Internal Medicine of Guangdong Cardiovascular Institute,Guangzhou 510010,China)

机构地区：[1]华南理工大学医学院,广东广州510006 [2]华南理工大学生物科学与工程学院,广东广州510006 [3]广东省人民医院(广东省医学科学院)医学研究部临床药理重点实验室,广东广州510080 [4]广东省心血管病研究所心血管内科,广东广州510100

出　　处：《中国病理生理杂志》2022年第4期577-583,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.82170415);广东省自然科学基金资助项目(No.2021A1515011551);广州市科技计划项目(No.202102080385);心血管疾病研究专项项目(No.2020XXG001);高水平医院建设项目(No.DFJH201925)。

摘　　要：目的:探讨钙感受器基质相互作用分子1(STIM1)参与主动脉平滑肌细胞衰老的作用研究。方法:采用过氧化氢(H_(2)O_(2))诱导人主动脉平滑肌细胞建立细胞衰老模型,并用衰老相关β-半乳糖苷酶(SA-β-Gal)染色检测细胞衰老变化;利用Cre-loxP基因重组技术,制备平滑肌特异性STIM1基因敲除(sm-STIM1-KO)小鼠;体内及体外干预STIM1表达,观察主动脉平滑肌细胞衰老变化。结果:在200μmol/L H_(2)O_(2)诱导的主动脉平滑肌细胞衰老模型中,SA-β-Gal染色细胞阳性率显著升高,同时钙库操纵性钙通道相关蛋白STIM1和ORAI1的表达显著下调(P<0.05)。另外,成功构建了sm-STIM1-KO小鼠,该小鼠主动脉的衰老相关蛋白p53、p21及p16表达上调(P<0.05)。用STIM1 siRNA敲减主动脉平滑肌细胞STIM1,衰老相关蛋白p21和p16显著上调(P<0.05)。在衰老细胞模型中,用腺病毒过表达STIM1则衰老相关蛋白p21和p16显著下调(P<0.01)。结论:钙感受器STIM1下调可加速血管平滑肌细胞的衰老,并参与血管老化的进展。AIM:To invest the effect of calcium sensor stromal interaction molecule 1(STIM1)on senes⁃cence of aortic smooth muscle cells.METHODS:Human aortic smooth muscle cells were treated with hydrogen peroxide(H_(2)O_(2))to establish the cellular senescence model.The senescent cells were detected by senescence-associatedβ-galacto⁃sidase(SA-β-Gal)staining.Smooth muscle-specific STIM1 gene knockout(sm-STIM1-KO)mice were constructed by Cre-loxP gene recombination technique.Effect of STIM1 on the senescence of aortic smooth muscle cells was observed after in⁃tervening STIM1 expression in vivo and in vitro.RESULTS:Treatment with H_(2)O_(2)(200μmol/L)induced senescence of human aortic smooth muscle cells,as indicated by increased SA-β-Gal positive rate.The expression of store-operated cal⁃cium channel-related proteins STIM1 and ORAI1 was significantly down-regulated by H_(2)O_(2)(P<0.05).The expression of senescence-related proteins p53,p21 and p16 were up-regulated in the aorta of sm-STIM1-KO mice compared with control mice(P<0.05).Senescence-related proteins p21 and p16 were significantly up-regulated in aortic smooth muscle cells with siRNA-mediated STIM1 knockdown(P<0.05).Over-expression of STIM1 by adenovirus significantly down-regulated p21 and p16 in the H_(2)O_(2)-induced senescent cells(P<0.01).CONCLUSION:Down-regulation of STIM1 accelerates the senescence of vascular smooth muscle cells and participates in the progression of vascular senescence.

关 键 词：基质相互作用分子1 基因敲除 细胞衰老 主动脉平滑肌细胞 钙库操纵性钙通道 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R363.2[医药卫生—基础医学]