Sirt3抑制血管紧张素Ⅱ诱导的小鼠血管平滑肌细胞增殖  

Inhibitory effect of Sirt3 on proliferation of vascular smooth muscle cells induced by angiotensin Ⅱ

作　　者：吴新宁[1] 卜培莉[1] 刘军妮[1] 赵立祥[1] 王希[1] 李娜[1] 

机构地区：[1]教育部和卫生部心血管重构与功能研究重点实验室,山东大学齐鲁医院心内科,山东济南250012

出　　处：《细胞与分子免疫学杂志》2013年第3期237-241,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81070076;81170135);国家重点基础研究发展计划(973)(2012CB722406);山东大学研究生自主创新基金(21300070613085)

摘　　要：目的检测小鼠血管平滑肌细胞(VSMCs)中Sirtuin3(Sirt3)的表达,探讨Sirt3在血管紧张素Ⅱ(AngⅡ)诱导小鼠VSMCs增殖中的作用。方法用RT-PCR、Western blot法检测细胞中是否有Sirt3基因和蛋白表达;用不同浓度AngⅡ(10-7mol/L、10-6mol/L、10-5mol/L)分别刺激细胞,用RT-PCR、Western blot法检测AngⅡ对Sirt3表达的影响;用Sirt3 siRNA转染干扰Sirt3mRNA水平后5-乙炔基-2'脱氧尿嘧啶核苷(Edu)试剂盒检测细胞增殖水平。结果 Western blot法、RT-PCR结果均显示正常C57小鼠VSMCs中有一定量Sirt3蛋白和mRNA表达;10-7mol/L、10-6mol/L、10-5mol/L AngⅡ刺激细胞后Sirt3 mR-NA水平及蛋白表达量均明显升高(P<0.01),其中10-6mol/L组比10-7mol/L组和10-5mol/L组升高更明显,差异有统计学意义(P<0.05);Sirt3敲减组细胞增殖较对照组明显增加(P<0.01),Sirt3沉默+AngⅡ(10-6mol/L)组较AngⅡ组细胞增殖明显增加(P<0.01)。结论小鼠VSMCs中有一定量的Sirt3表达,AngⅡ刺激可使Sirt3 mRNA水平和蛋白表达量明显升高;Sirt3可抑制AngⅡ诱导的VSMCs增殖。Objective To detect the expression of Sirtuin3 （SirL3） in mice vascular smooth muscle cells （VSMCs） and explore the effect of Sirl3 on VSMCs proliferation induced by angiotensin Ⅱ （ Ang Ⅱ ）. Methods The mRNA and protein expressions of Sirt3 in wild C57 mice VSMCs were assessed by RT-PCR and Western blotting, respectively. After the cells were exposed to various concentrations of Ang Ⅱ （ 10-7, 10-6, 10-5 mol/L）, the mRNA and protein expressions of Sirl3 were assessed again in the same way. The effect of Sirl3 on cell proliferation was observed with Edu （5-ethynyl -2＇-deoxyuri- dine） kit after Sirl3 silencing by small interference RNA. Results Sirt3 was stably expressed in C57 mice VSMCs as revealed by Western blotting and RT-PCR, respectively. A significant increase was found in the mRNA and protein expressions of Sirt3 （P 〈 0.01 ） after the stimulation of all the three different concentrations of Ang Ⅱ, particularly the 10-6 mol/L group with the highest increase of Sirt3 expression, and there was a significant difference between the 10-6 moVL group and the other groups （ P〈0.05）. The rate of cell proliferation in Sirt3 SiRNA group increased obviously compared to the control group （ P〈 0.01 ）, and the rate in the Sirl3 SiRNA ＋ Ang Ⅱ group was also elevated significantly compared to the Ang Ⅱgroup （ P 〈 0.01 ）. Conclusion The expression of Sirt3 is stable in VSMCs; Ang Ⅱ can elevate the mRNA and protein expression of Sift3 and promote the cell proliferation, what＇s more, Sirt3 silencing would further increase the cell proliferation.

关 键 词：沉默信号调节因子3 血管紧张素Ⅱ 血管平滑肌细胞 增殖 

分 类 号：R392.12[医药卫生—免疫学]