普伐他汀预处理对香烟提取物诱导的小鼠血管平滑肌细胞凋亡的影响及可能机制研究  

Impact of Pravastatin Pretreatment on Cigarette Smoke Extract-induced Apoptosis of Vascular Smooth Muscle Cells in Mice and Its Mechanism

作　　者：廖思聪 祁春雷 王大新[2] LIAO Si-cong;QI Chun-lei;WANG Da-xin(Department of Cardiology, the Second Xiangya Hospital of Central South University, Changsha 410011, China;Department of Cardiology, the People＇ s Hospital Northern Jiangsu, Yangzhou 225001, China)

机构地区：[1]中南大学湘雅二医院心内科,湖南省长沙市410011 [2]江苏省苏北人民医院心内科,江苏省扬州市225001

出　　处：《实用心脑肺血管病杂志》2018年第3期44-50,共7页Practical Journal of Cardiac Cerebral Pneumal and Vascular Disease

基　　金：国家重大基础研究项目"973"计划项目(2007CB936104);江苏省"六大人才高峰"项目(2014-SWYY-052)

摘　　要：目的探讨普伐他汀预处理对香烟提取物(CSE)诱导的小鼠血管平滑肌细胞(VSMCs)凋亡的影响及可能机制。方法 2017年6月—2018年1月,采用负压吸引装置抽吸香烟烟雾并经过滤除菌制备CSE原液,取10代以内生长状态良好的对数期VSMCs进行实验。(1)将实验细胞分为不同浓度CSE诱导组,其中空白组使用不含CSE的DMEM培养基培养12 h,低浓度组、中浓度组、高浓度组分别于含1%、5%、10%CSE的DMEM培养基培养12 h。(2)将实验细胞分为不同剂量普伐他汀预处理组,其中对照组使用不含普伐他汀的DMEM培养基培养12 h;低剂量组、中剂量组、高剂量组分别给予1μM、10μM、100μM普伐他汀预处理2 h,然后使用含5%CSE的DMEM培养基培养12 h;部分实验设置阳性对照组,阳性对照组给予4-苯丁酸(4-PBA)2 m M预处理2 h,然后使用含5%CSE的DMEM培养基培养12 h。采用流式细胞术检测细胞凋亡率,采用实时荧光定量聚合酶链反应检测Bcl-2、Bax、GRP78、Chop m RNA相对表达量,采用Western Blot检测Bcl-2、Bax、GRP78、Chop蛋白相对表达量。结果 (1)低浓度组、中浓度组和高浓度组细胞凋亡率高于空白组,中浓度组和高浓度组细胞凋亡率高于低浓度组,高浓度组细胞凋亡率高于中浓度组(P<0.05)。(2)低浓度组、中浓度组和高浓度组Bcl-2 m RNA和蛋白相对表达量低于空白组,中浓度组和高浓度组Bcl-2 m RNA和蛋白相对表达量低于低浓度组,高浓度组Bcl-2蛋白相对表达量低于中浓度组(P<0.05);低浓度组、中浓度组和高浓度组Bax m RNA和蛋白相对表达量高于空白组,中浓度组和高浓度组Bax m RNA和蛋白相对表达量高于低浓度组,高浓度组Bax m RNA相对表达量高于中浓度组(P<0.05)。(3)低浓度组、中浓度组、高浓度组GRP78、Chop m RNA和蛋白相对表达量高于空白组,中浓度组、高浓度组GRP78、Chop m RNA和蛋白相对表达量高于低浓度组,高浓度组GRP78 m RNA、Chop蛋白相对�Objective To investigate the impact of pravastatin pretreatment on cigarette smoke extract（CSE）-induced apoptosis of vascular smooth muscle cells（VSMCs） in mice and its mechanism. Methods This experiment was carried out from June 2017 to January 2018. Negative pressure suction device was used to collect cigarette smoke, and CSE stoste was prepared by filtration sterilization, log phase VSMCs（within 10 cell propagations） with good growing status were selected for the experiment.（1） VSMCs were divided into blank group（cultivated without CSE）, low-concentration group（cultivated with 1% CSE）, moderate-concentration group（cultivated with 5% CSE） and high-concentration group（cultivated with 10% CSE） according to the concentration of CSE, and then were cultivated for 12 hours on DMEM medium.（2） VSMCs were divided into control group（without pravastatin pretreatment）, low-dose group（with 1 μM pravastatin pretreatment for 2 hours）, moderate-dose group（with 10 μM pravastatin pretreatment for 2 hours） and high-dose group（with 100 μM pravastatin pretreatment for 2 hours） according to the concentration of CSE, and then were cultivated for 12 hours on DMEM medium with 5% CSE. Meanwhile some experiments set positive control group（with 2 m M 4-PBA pretreatment for 2 hours and then were cultivated for 12 hours on DMEM medium with 5% CSE）. Flow cytometry was used to detect the apoptosis rate, real-time fluorescence quantitative PCR was used to detect the relative expression quantity of m RNA of Bcl-2, Bax, GRP78 and Chop, and Western Blot was used to detect the relative expression quantity of protein of Bcl-2, Bax, GRP78 and Chop. Results（1） Apoptosis rate in low-concentration group, moderate-concentration group and high-concentration group was statistically significantly higher than that in blank group, respectively, apoptosis rate in moderate-concentration group and high-concentration group was statistically significantly higher than that in low-concentration group, r

关 键 词：动脉粥样硬化 肌细胞 平滑肌 细胞凋亡 普伐他汀 香烟提取物 

分 类 号：R543.5[医药卫生—心血管疾病]