阿司匹林抑制血小板源性生長因子BB引起的主動(dòng)脈血管平滑肌細(xì)胞增殖和遷移研究
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本文關(guān)鍵詞:阿司匹林抑制血小板源性生長因子BB引起的主動(dòng)脈血管平滑肌細(xì)胞增殖和遷移研究 出處:《中國全科醫(yī)學(xué)》2016年36期 論文類型:期刊論文
更多相關(guān)文章: 阿司匹林 血小板源性生長因子 主動(dòng)脈 肌細(xì)胞 平滑肌 細(xì)胞增殖 細(xì)胞運(yùn)動(dòng)
【摘要】:目的探討阿司匹林對由血小板源性生長因子BB(PDGF-BB)引起的主動(dòng)脈血管平滑肌細(xì)胞增殖和遷移的影響及其機(jī)制。方法 2014年8月—2015年9月,選取人主動(dòng)脈血管平滑肌細(xì)胞株(T/G HA-VSMC),分為對照組(不添加任何試劑)、PDGF-BB組(添加10 ng/ml PDGF-BB)、阿司匹林組(添加5 mmol/L阿司匹林)、PDGF-BB+阿司匹林組(添加10 ng/ml PDGF-BB和5 mmol/L阿司匹林),采用CCK-8法檢測4組不同培養(yǎng)時(shí)間(0、24、48、72 h)T/G HA-VSMC增殖吸光度(OD值),Western blotting法檢測p21waf1、p27kip1相對表達(dá)量,流式細(xì)胞儀檢測細(xì)胞周期,細(xì)胞劃痕實(shí)驗(yàn)檢測細(xì)胞遷移率,Western blotting法檢測基質(zhì)金屬蛋白酶1(MMP-1)相對表達(dá)量。結(jié)果培養(yǎng)0 h時(shí),4組T/G HA-VSMC增殖OD值比較,差異無統(tǒng)計(jì)學(xué)意義(P0.05)。培養(yǎng)24、48、72 h時(shí),PDGF-BB組T/G HA-VSMC增殖OD值較對照組升高,阿司匹林組T/G HA-VSMC增殖OD值較對照組降低(P0.05);培養(yǎng)48、72 h時(shí),PDGF-BB+阿司匹林組T/G HA-VSMC增殖OD值較對照組降低(P0.05);培養(yǎng)24、48、72 h時(shí),阿司匹林組和PDGF-BB+阿司匹林組T/G HA-VSMC增殖OD值較PDGF-BB組降低(P0.05);培養(yǎng)24、48、72 h時(shí),PDGF-BB+阿司匹林組T/G HA-VSMC增殖OD值較阿司匹林組升高(P0.05)。PDGF-BB組培養(yǎng)24、48 h時(shí)p21~(waf1)、p27~(kip1)相對表達(dá)量較培養(yǎng)0 h時(shí)升高(P0.05);培養(yǎng)72 h時(shí)p21waf1、p27kip1相對表達(dá)量較培養(yǎng)0、24、48 h時(shí)降低(P0.05)。培養(yǎng)24 h時(shí),PDGF-BB組p21~(waf1)、p27~(kip1)相對表達(dá)量較對照組、阿司匹林組和PDGF-BB+阿司匹林組降低(P0.05)。PDGF-BB組G0/G1期細(xì)胞所占比例較對照組、阿司匹林組和PDGF-BB+阿司匹林組降低,S期細(xì)胞所占比例較對照組、阿司匹林組和PDGF-BB+阿司匹林組升高(P0.05)。細(xì)胞劃痕實(shí)驗(yàn)結(jié)果顯示,培養(yǎng)24 h時(shí),PDGF-BB組細(xì)胞遷移率較對照組、阿司匹林組和PDGF-BB+阿司匹林組降低(P0.05)。PDGF-BB組MMP-1相對表達(dá)量較對照組、阿司匹林組和PDGF-BB+阿司匹林組升高(P0.05);阿司匹林組和PDGF-BB+阿司匹林組MMP-1相對表達(dá)量較對照組降低(P0.05)。結(jié)論阿司匹林通過上調(diào)p21~(waf1)、p27~(kip1),下調(diào)MMP-1而抑制由PDGF-BB引起的主動(dòng)脈血管平滑肌細(xì)胞的增殖和遷移。
[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.
【作者單位】: 貴州省貴陽市第一人民醫(yī)院心血管內(nèi)科;第三軍醫(yī)大學(xué)附屬新橋醫(yī)院心血管內(nèi)科;
【基金】:貴州省科技廳社會(huì)發(fā)展資助項(xiàng)目(黔科合SY[2010]3087號(hào))
【分類號(hào)】:R96
【正文快照】: 血小板源性生長因子(platelet derived growth factor,PDGF)最初在血清和血小板中被發(fā)現(xiàn),能在體外誘導(dǎo)血管平滑肌細(xì)胞(VSMC)和成纖維細(xì)胞的分裂,因此被認(rèn)為是一種分裂素[1-2]。PDGF有5種二聚體形式,分別為:PDGF-AA、PDGF-BB、PDGF-CC、PDGF-DD和PDGF-AB,其中PDGF-BB在促進(jìn)細(xì)胞
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