阿司匹林抑制血小板源性生长因子BB引起的主动脉血管平滑肌细胞增殖和迁移研究  

Aspirin Inhibition of PDGF-BB- induced Proliferation and Migration of Vascular Smooth Muscle Cells

作　　者：朱晋坤[1] 毛华[1] 尹扬光[2] 董文[1] 杜峰[1] 黎姗姗[1] 邓梦扬[2] 

机构地区：[1]贵州省贵阳市第一人民医院心血管内科,550002 [2]第三军医大学附属新桥医院心血管内科

出　　处：《中国全科医学》2016年第36期4467-4473,共7页Chinese General Practice

基　　金：贵州省科技厅社会发展资助项目(黔科合SY[2010]3087号)

摘　　要：目的探讨阿司匹林对由血小板源性生长因子BB(PDGF-BB)引起的主动脉血管平滑肌细胞增殖和迁移的影响及其机制。方法 2014年8月—2015年9月,选取人主动脉血管平滑肌细胞株(T/G HA-VSMC),分为对照组(不添加任何试剂)、PDGF-BB组(添加10 ng/ml PDGF-BB)、阿司匹林组(添加5 mmol/L阿司匹林)、PDGF-BB+阿司匹林组(添加10 ng/ml PDGF-BB和5 mmol/L阿司匹林),采用CCK-8法检测4组不同培养时间(0、24、48、72 h)T/G HA-VSMC增殖吸光度(OD值),Western blotting法检测p21waf1、p27kip1相对表达量,流式细胞仪检测细胞周期,细胞划痕实验检测细胞迁移率,Western blotting法检测基质金属蛋白酶1(MMP-1)相对表达量。结果培养0 h时,4组T/G HA-VSMC增殖OD值比较,差异无统计学意义(P>0.05)。培养24、48、72 h时,PDGF-BB组T/G HA-VSMC增殖OD值较对照组升高,阿司匹林组T/G HA-VSMC增殖OD值较对照组降低(P<0.05);培养48、72 h时,PDGF-BB+阿司匹林组T/G HA-VSMC增殖OD值较对照组降低(P<0.05);培养24、48、72 h时,阿司匹林组和PDGF-BB+阿司匹林组T/G HA-VSMC增殖OD值较PDGF-BB组降低(P<0.05);培养24、48、72 h时,PDGF-BB+阿司匹林组T/G HA-VSMC增殖OD值较阿司匹林组升高(P<0.05)。PDGF-BB组培养24、48 h时p21^(waf1)、p27^(kip1)相对表达量较培养0 h时升高(P<0.05);培养72 h时p21waf1、p27kip1相对表达量较培养0、24、48 h时降低(P<0.05)。培养24 h时,PDGF-BB组p21^(waf1)、p27^(kip1)相对表达量较对照组、阿司匹林组和PDGF-BB+阿司匹林组降低(P<0.05)。PDGF-BB组G0/G1期细胞所占比例较对照组、阿司匹林组和PDGF-BB+阿司匹林组降低,S期细胞所占比例较对照组、阿司匹林组和PDGF-BB+阿司匹林组升高(P<0.05)。细胞划痕实验结果显示,培养24 h时,PDGF-BB组细胞迁移率较对照组、阿司匹林组和PDGF-BB+阿司匹林组降低(P<0.05)。PDGF-BB组MMP-1相对表达量较对照组、阿司匹林组和PDGF-BB+阿司匹林组升高(P<0.05);阿司匹林组和PDGF-BB+Objective To explore the influence and mechanism of aspirin on proliferation and migration of aortic vascular smooth muscle cells caused by platelet- derived growth factor BB（ PDGF-BB）. Methods From August 2014 to September 2015,aortic vascular smooth muscle cell strains（ T/G HA- VSMC） were selected and divided into four groups：control group（ no other reagent）,PDGF-BB group（ with 10 ng/ml PDGF-BB）,aspirin group（ with 5 mmol/L aspirin）,and PDGF-BB ＋ aspirin group（ with 10 ng/ml PDGF-BB and 5 mmol/L aspirin）. OD value of T/G HA- VSMC proliferation at different cultivation times（ 0,24,48,and 72 h） was tested by CCK- 8 method,and relative expression quantity of p21^（waf1） and p27^（kip1） was tested by Western blotting. Cell cycles were tested by flow cytometry,and cell migration was tested by wound-healing assay. Relative expression quantity of matrix metalloproteinase 1（ MMP-1） in four groups was tested by Western blotting.Results When cultivated at 0 h,differences of OD value of T/G HA- VSMC proliferation in four groups showed no statistical significance（ P 〉0. 05）. When cultivated at 24,48,and 72 h,OD value of T/G HA- VSMC proliferation in PDGF-BB group was higher than that in control group（ P 〈0. 05）. In aspirin group,OD value of T/G HA- VSMC proliferation was lower than that in control group（ P 〈0. 05）; when cultivated at 48 and 72 h,OD value of T/G HA- VSMC proliferation in PDGF-BB ＋aspirin group was lower than that in control group（ P 〈0. 05）; when cultivated at 24,48, and 72 h, OD value of T/G HA- VSMC proliferation in aspirin group and PDGF-BB ＋ aspirin group was lower than that in PDGF-BB group（ P 〈0. 05）;when cultivated at 24,48,and 72 h,OD value of T/G HA- VSMC proliferation in PDGF-BB ＋ aspirin group was higher than that in aspirin group（ P 〈0. 05）. Relative expression quantities of p21waf1 and p27kip1 in PDGF-BB group at 24 and 48 h were higher than those at 0 h（ P 〈0. 05）; when cultivated at 72 h,relative express

关 键 词：阿司匹林 血小板源性生长因子 主动脉 肌细胞 平滑肌 细胞增殖 细胞运动 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]