高钙磷激活DNA损伤应答诱导人主动脉平滑肌细胞早衰  

DNA damage response activated by high calcium and phosphorus induces premature aging of human aortic smooth muscle cells

作　　者：范志娟[1] 武玉晶 田亚琼[1] 刘爽[1] 张蝶 刘树业[1] FAN Zhijuan;WU Yujing;TIAN Yaqiong;LIU Shuang;ZHANG Die;LIU Shuye(Department of Clinical Laboratory,the Third Central Hospital of Tianjin,Tianjin Key Laboratory of Extracorporeal Life Support for Critical Diseases,Artificial Cell Engineering Technology Research Center,Tianjin Institute of Hepatobiliary Disease,Tianjin 300170,China)

机构地区：[1]天津市第三中心医院检验科天津市重症疾病体外生命支持重点实验室天津市人工细胞工程技术研究中心天津市肝胆疾病研究所,天津300170

出　　处：《基础医学与临床》2023年第8期1234-1240,共7页Basic and Clinical Medicine

基　　金：天津市医学重点学科(TJYXZDXK-047A);天津市卫生健康委科技基金(KJ20151)。

摘　　要：目的探讨DNA损伤应答(DDR)通路调控人主动脉平滑肌细胞(HASMCs)钙化机制。方法将HASMCs分为对照组、模型组、ATM干预组、PARP干预组,培养12 d。茜素红-S染色法定性和邻-甲酚酞法定量检测细胞钙化;Western blot检测组蛋白γH2AX磷酸化、p16和p21、ATM上Ser1981的磷酸化水平;β-半乳糖苷酶染色检测细胞早衰;qPCR检测p16和p21 mRNA水平。8-羟基-2′-脱氧鸟苷(8-OHDG)检测氧化应激水平,ELISA方法检测IL-6、IL-8水平。结果模型组较对照组钙化明显,8-OHDG、组蛋白γH2AX磷酸化、β-半乳糖苷酶染色、p16的mRNA和蛋白、p21 mRNA、IL6和IL8、ATM磷酸化等指标有显著变化(P<0.05),ATM和PARP干预组可以缓解模型组的变化。结论高钙磷环境刺激HASMCs产生持续DNA损伤,触发ATM磷酸化并激活p16蛋白表达,诱导细胞早衰导致钙化。Objective To investigate the mechanism of DNA damage response(DDR)pathway regulating calcification in human aortic smooth muscle cells(HASMCs).Methods The HASMCs were divided into the control group,model group,ATM treatment group,and PARP treatment group,and they were cultured for 12 days.Cell calcification was measured by Alizarin red staining andσ-Cresolphthalein;phosphorylation levels of histoneγH2AX,protein levels of p16 and p21,and phosphorylation levels of ATM on Ser1981 were tested by Western blot,premature cell senescence byβ-galactosidase staining;and p16 and p21 mRNA by qPCR.The level of oxidative stress was measured by 8-hydroxy-2′-deoxyguanosine(8-OHDG),and the level of IL-6 and IL-8 was measured by ELISA kit.Results The calcification was evident in the model group as compared with that in control group.There were significant changes in 8-OHDG,histoneγH2AX phosphorylation,β-galactosidase staining,mRNA and protein of p16,p21 mRNA,release of IL 6 and IL 8 and ATM phosphorylation(P<0.05).The changes in the model group alleviated by ATM and PARP treatment.Conclusions High calcium and phosphorus environment stimulates HASMCs to produce sustained DNA damage,triggers ATM phosphorylation,activates p16 protein expression,and induces premature cell senescence causing cell death and resulted in calcification.

关 键 词：Ca^(2+)/P DNA损伤应答 细胞早衰 人主动脉平滑肌细胞 血管钙化 

分 类 号：R543.5[医药卫生—心血管疾病]