SIRT1上调大鼠血管平滑肌细胞P27^(Kip1)的表达  

Upregulation of P27^(Kip1) expression by SIRT1 in rat vascular smooth muscle cells

作　　者：鲁云彪[1] 陈厚早[1] 张冉[1] 郑伟[1] 李莉[1] 张庆军[1] 刘德培[1] 

机构地区：[1]中国医学科学院基础医学研究所北京协和医学院基础学院医学分子生物学国家重点实验室,北京100005

出　　处：《基础医学与临床》2010年第7期714-720,共7页Basic and Clinical Medicine

基　　金：国家重点基础研究发展计划(973)(2006CB503801);国家高技术研究发展计划(863)(2006AA02A406);国家重点实验室专项经费(2060204)

摘　　要：目的研究Ⅲ类去乙酰化酶SIRT1在血管平滑肌中对P27Kip1表达的影响及其可能的机制。方法用H2O2和oxLDL刺激大鼠平滑肌细胞A7r5,Western blot检测平滑肌细胞内源性SIRT1的表达变化;在动物血管损伤模型中,Western blot检测血管损伤处平滑肌的SIRT1表达水平。用SIRT1重组腺病毒感染A7r5细胞和原代大鼠平滑肌细胞,Western blot观察SIRT1过表达引起的P27表达的改变;用SIRT1抑制剂NAM处理平滑肌细胞并观察P27的表达改变;用免疫共沉淀技术探寻SIRT1上调P27基因表达可能的分子机制。结果在H2O2和oxLDL氧化应激刺激的A7r5细胞中,内源性SIRT1的表达随作用时间逐渐增加;动物血管损伤模型中,内源性SIRT1的表达明显上调;过表达SIRT1能够在10%血清刺激的早期上调P27表达,而SIRT1抑制剂NAM则能够减少P27表达;在原代平滑肌细胞中,腺病毒介导的SIRT1过表达同样能够显著上调P27的表达。在血管平滑肌细胞中检测到SIRT1能够与FOXO3a相互作用。结论 SIRT1在血管平滑肌细胞中上调P27的表达。Objective To explore the impact of SIRT1 on P27Kip1 expression in vascular smooth muscle cells（VSMCs） and to investigate the possible mechanism.Methods A7r5 VSMCs were treated wtih H2O2 or oxLDL,then SIRT1 expression was detected by Western blot.Vascular injury models were also used to detect SIRT1 expression by Western blot.Primary rat VSMCs were infected with wild-type SIRT1 adenovirues and examined the expression of P27 by Western blot.Then we treated quiescent cells with nicotinamide,an inhibitor of the deacetylase activity of SIRT1,then detected potential change of P27 expression.We also detected the interaction between SIRT1 and FOXO3a in VSMCs by co-immunoprecipitation.Results Endogenous SIRT1 was upregulated in a time-dependent manner in H2O2 or oxLDL treated A7r5 cells,and also upregulated in vascular injury models.SIRT1 overexpression significantly upregulated P27 expression under 10%FBS stimulation.NAM was found to inhibit P27 expression in a dose-dependent manner.The interaction between SIRT1 and FOXO3a was detectable in smooth muscle cells.Conclusion SIRT1 upregulates P27 expression in vascular smooth muscle cells.

关 键 词：SIRT1 P27KIP1 血管平滑肌细胞 

分 类 号：R543[医药卫生—心血管疾病]