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

发布时间：2018-01-18 10:19

本文关键词：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　出处：《中国全科医学》2016年36期 　论文类型：期刊论文

【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(P0.05)。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(P0.05);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(P0.05);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(P0.05);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(P0.05)。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(P0.05);培养72 h时p21waf1、p27kip1相对表达量较培养0、24、48 h时降低(P0.05)。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(P0.05)。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(P0.05)。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(P0.05)。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(P0.05);阿司匹林组和PDGF-BB+阿司匹林组MMP-1相对表达量较对照组降低(P0.05)。结论阿司匹林通过上调p21~(waf1)、p27~(kip1),下调MMP-1而抑制由PDGF-BB引起的主动脉血管平滑肌细胞的增殖和迁移。
[Abstract]:Objective to investigate the effect of aspirin on the platelet derived growth factor BB (PDGF-BB) on proliferation and migration of vascular smooth muscle cells induced and its mechanism. Methods from August 2014 to September 2015, from human aortic vascular smooth muscle cells (T/G HA-VSMC), divided into control group (without any reagent), PDGF-BB group (adding 10 ng/ml PDGF-BB), aspirin group (adding 5 mmol/L, PDGF-BB+ aspirin group (aspirin) containing 10 ng/ml PDGF-BB and 5 mmol/L aspirin), CCK-8 method was used to detect 4 groups of different culture time (0,24,48,72 h) T/G HA-VSMC absorbance (OD), proliferation detection p21WAF1 Western blotting method. The relative expression of p27kip1, cell cycle was analyzed by flow cytometry cell scratch assay, cell migration rate, detection of matrix metalloproteinase Western (MMP-1) blotting 1 phase of expression. Results cultured for 0 h, 4 T/G HA-VSMC group Colonization od comparison, no statistically significant difference (P0.05). 24,48,72 h, PDGF-BB T/G group, the proliferation of HA-VSMC was higher than the control group, aspirin group T/G HA-VSMC proliferation OD values were lower than the control group (P0.05); culture of 48,72 h, PDGF-BB+ T/G HA-VSMC ressor od aspirin group was lower than control group (P0.05); culture 24,48,72 h, aspirin group and aspirin group decreased PDGF-BB+ T/G HA-VSMC proliferation OD value compared with the PDGF-BB group (P0.05); culture of 24,48,72 h, PDGF-BB+ T/G HA-VSMC aspirin group od proliferation compared with the aspirin group increased (P0.05).PDGF-BB 24,48 h p21~ group (WAF1), p27~ (kip1) relative expression was cultured for 0 h increased (P0.05); 72 h culture p21WAF1, p27kip1 relative expression amount decrease when cultured 0,24,48 H (P0.05). After 24 h of culture, PDGF-BB p21~ group (WAF1), p27~ (kip1) relative expression compared with the control group, aspirin group and PDGF-BB+. Reduce the aspirin group (P0.05) for.PDGF-BB group the proportion of cells in G0/G1 phase decreased compared with the control group, aspirin group and aspirin group PDGF-BB+, the proportion of cells at S phase compared with the control group, aspirin group and aspirin group increased PDGF-BB+ (P0.05). Cell scratch test results showed that 24 h culture, PDGF-BB cell migration lower rate than the control group, aspirin group and aspirin group (P0.05 PDGF-BB+) group.PDGF-BB MMP-1 relative expression compared with the control group, aspirin group and aspirin group increased PDGF-BB+ (P0.05); aspirin group and aspirin group PDGF-BB+ MMP-1 relative expression was lower than control group (P0.05). Conclusion aspirin through upregulation of p21~ (WAF1), p27~ (kip1), down-regulation of MMP-1 and inhibit the proliferation and migration of PDGF-BB induced vascular smooth muscle cells.

【作者单位】：贵州省贵阳市第一人民医院心血管内科;第三军医大学附属新桥医院心血管内科;
【基金】：贵州省科技厅社会发展资助项目(黔科合SY[2010]3087号)
【分类号】：R96
【正文快照】： 血小板源性生长因子(platelet derived growth factor,PDGF)最初在血清和血小板中被发现,能在体外诱导血管平滑肌细胞(VSMC)和成纤维细胞的分裂,因此被认为是一种分裂素[1-2]。PDGF有5种二聚体形式,分别为:PDGF-AA、PDGF-BB、PDGF-CC、PDGF-DD和PDGF-AB,其中PDGF-BB在促进细胞

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