【摘要】：目的探討纖溶酶原激活物抑制物-1(PAI-1)對(duì)氣道平滑肌增殖的調(diào)控作用,以及對(duì)細(xì)胞外調(diào)節(jié)蛋白激酶(ERK)表達(dá)的影響。方法體外培養(yǎng)小鼠氣道平滑肌細(xì)胞(ASMCs)并分為7組:空白對(duì)照組、PAI-15μg/L組、PAI-1 10μg/L組、PAI-1 20μg/L組、PAI-1 40μg/L組、PAI-1 80μg/L組、PAI-1 100μg/L組。放入37℃培養(yǎng)箱分別培養(yǎng)12、24和48 h,采用CCK-8檢測(cè)ASMCs的A值,計(jì)算增殖率。以增殖率最高的實(shí)驗(yàn)組的濃度和作用時(shí)間作為PAI-1作用最適濃度和最適作用時(shí)間,進(jìn)一步分6組:A組(空白對(duì)照);B組(PAI-1 20μg/L);C組(PAI-1 40μg/L);D組(PAI-1 80μg/L);E組(PAI-1 100μg/L);F組(PAI-1最適濃度+ERK通道抑制劑PD98059 10μmol/L),用CCK-8檢測(cè)ASMCs的增殖,Western blot檢測(cè)ERK蛋白的表達(dá),實(shí)時(shí)熒光定量聚合酶鏈反應(yīng)檢測(cè)ERK m RNA的表達(dá)。結(jié)果 5~100μg/L PAI-1作用ASMCs 12、24和48 h后細(xì)胞增殖率結(jié)果顯示,在相同濃度下,不同時(shí)間各組比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05),以48 h時(shí)ASMCs增殖率最高;在相同時(shí)間下,不同濃度各組比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05),80μg/L PAI-1的ASMCs增殖率最高;故選取48 h為最適作用時(shí)間,80μg/L為最適濃度進(jìn)行后續(xù)實(shí)驗(yàn)。再次分組的實(shí)驗(yàn)結(jié)果表明,B、C、D、E組的ERK磷酸化水平和ERK m RNA的相對(duì)表達(dá)量與A組比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05),B、C、D、E組增高;B、C、D、E組兩兩比較結(jié)果顯示,除B組與E組比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)外,其余各組兩兩比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05),當(dāng)PAI-1濃度波動(dòng)于20~80μg/L時(shí),ERK磷酸化水平和ERK m RNA相對(duì)表達(dá)量隨濃度升高而增高,80μg/L后則不再增高;加入PD98059的F組與D組比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05),ERK磷酸化水平和ERK m RNA表達(dá)降低。結(jié)論外源性PAI-1可以通過(guò)促進(jìn)ERK通路的表達(dá),進(jìn)而促進(jìn)ASMCs的增殖。
[Abstract]:Objective to investigate the regulatory effect of plasmin activating inhibitor-1 (PAI-1) on the proliferation of airway smooth muscle and the expression of extracellular regulated protein kinase (ERK). Methods (ASMCs) of mouse airway smooth muscle cells were cultured in vitro and divided into 7 groups: blank control group, PAI- 15 渭 g / L group, PAI-1 10 渭 g / L group, PAI-1 20 渭 g / L group, PAI-1 40 渭 g / L group, PAI-1 80 渭 g / L group, PAI-1 100 渭 g / L group. The A value of ASMCs was detected by CCK-8 in 37 鈩，

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