白藜三醇抑制血管平滑肌细胞增殖及其相关机制  

Resveratrol inhibits the proliferation of vascular smooth muscle cells and related mechanisms

作　　者：朱启仁 李飞[2] 张梦雨 刘晓虹 李菲 李承宗 徐晤 张卓琦 王志荣 ZHU Qiren;LI Fei;ZHANG Mengyu;LIU Xiaohong;LI Fei;LI Chengzong;XU Wu;ZHANG Zhuoqi;WANG Zhirong(Department of Cardiology,the Affiliated Hospital of Xuzhou Medical University,Xuzhou,Jiangsu 221002,China;Department of Cardiology,Pizhou People＇s Hospital,Pizhou,Jiangsu 221300;Graduate School,Xuzhou Medical University,Xuzhou,Jiangsu 221004)

机构地区：[1]徐州医科大学附属医院心血管内科,江苏徐州221002 [2]邳州市人民医院心血管内科,江苏邳州221300 [3]徐州医科大学研究生学院,江苏徐州221004

出　　处：《徐州医科大学学报》2018年第7期453-458,共6页Journal of Xuzhou Medical University

基　　金：徐州市医学青年后备人才项目

摘　　要：目的探讨白藜三醇（RSV）对大鼠胸主动脉血管平滑肌细胞（VSMCs）增殖的调控及其相关机制。方法采用组织块贴壁法体外培养VSMCs，细胞分为3组：空白组（CON组）、FBS组（含20％胎牛血清的培养基）、RSV组。CCK-8法检测血管平滑肌细胞活性以确定RSV最佳作用浓度及时间，qPCR法检测各组细胞内mi-croRNAs的表达水平。为了证实RSV是否通过调控miR-21的表达发挥抗VSMCs增殖作用，除了上述3组，另增加miR-21过表达组（miR-21mimics组）和沉默组（miR-21inhibitor组）：miR-21mimics＋RSV组、miR-21inhibitor组、miR-21mimics组和miR-21转染阴性对照组（NC组），Westernblot检测各组人第10号染色体缺失的磷酸酶及张力蛋白同源的基因（PTEN）、增殖细胞核抗原（PCNA）蛋白的表达水平，流式细胞术（FCM）检测各组细胞周期明确细胞增殖状况。结果与CON组相比，FBS组miR-21表达明显上调（P〈0．05），RSV组miR-21表达明显下调（P〈0．05）。与CON组相比，FBS组和miR-21mimics组PTEN蛋白表达量下降（P〈0．05），而细胞明显增殖（P〈0．05）；RSV组和miR-21inhibitor组PTEN蛋白表达量升高（P〈0．05），PCNA蛋白表达下降，细胞增殖明显抑制（P〈0．05）。与miR-21mimics＋RSV组相比，miR-21mimics组PTEN蛋白表达量下降，而PCNA蛋白表达增多，细胞明显增殖（P〈0．05），RSV组则相反（P〈0．05）。结论RSV通过抑制miR-21表达而调控其下游靶基因这一途径在大鼠VSMCs中发挥抗增殖作用。Objective To investigate the regulation of resveratrol （RSV） on the proliferation of rat thoracic aorta vascular smooth muscle cells （VSMCs） and related mechanisms. Methods VSMCs were cultured in vitro using tissue explants. The cells were divided into three groups： a CON group, a FBS group （containing 20% FBS in the media） and an RSV group. The activities of vascular smooth muscle cells were measured by CCK - 8 assay to determine the optimal concentration of RSV and time. The levels of microRNAs in each group were examined by qPCR method. In order to confirm whether RSV inhibited the proliferation of VSMCs by regulating the expression of miR- 21, a miR- 21 overexpression group and a miR -21silencing group were added： a miR -21 mimics ＋ RSV group, a miR -21 inhibitor group, a miR -21 mimics group and a miR -21 negative control group （NC group）. The expression of phosphatase and tensin homologdeleted on chromosome ten （PTEN） and proliferating cell nuclear antigen （PCNA） was detected by Western blotting. The cell cycle of all group were examined by flow cytometry （FCM）. Results The expression of miR- 21 was significantly increased for the FBS groups and significantly decreased for the RSV group, compared with those in the CON group （P 〈0.05）. Compared with the CON group, the FBS and miR- 21 mimics groups showed decreased levels of PTEN （P 〈 0.05 ） and significant cell proliferation （ P 〈 0.05 ）, while increased PTEN levels, decreased PCNA amounts, and significantly inhibited cell proliferation were found in the RSV and miR - 21 inhibitor groups （ P 〈 0.05 ）. Compared with the miR- 21 mimics ＋ RSV group, the miR- 21 mimics group presented decreased PTEN levels, increased PCNA amounts, and significantly stimulated cell proliferation （ P 〈 0.05 ）, while the opposite changes were found in the RSV group （ P 〈 0.05）. Conclusions RSV exerts anti - proliferative effects on rat VSMCs by inhibiting miR - 21 expression and regulating its downstream target

关 键 词：血管平滑肌细胞 白藜三醇 MICRORNA-21 PTEN 增殖细胞核抗原 

分 类 号：R541.4[医药卫生—心血管疾病]