本文選題：心肌纖維化 + 活性氧　； 參考：《吉林大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)》2017年05期

【摘要】：目的:探討高血壓過(guò)程中心肌成纖維細(xì)胞(CFs)氧化應(yīng)激水平與中電導(dǎo)鈣激活鉀離子通道(KCa3.1)蛋白表達(dá)的關(guān)系,闡明KCa3.1在心肌纖維化中的作用及機(jī)制。方法:原代培養(yǎng)雄性AGT-REN雙轉(zhuǎn)基因高血壓小鼠(dTH)CFs,另培養(yǎng)同品系野生C57B6小鼠CFs作為對(duì)照。dTH小鼠CFs隨機(jī)分為高血壓組(dTH)和N-乙酰半胱氨酸組(NAC),dTH小鼠CFs給予不同濃度NAC干預(yù)24h。MTT法檢測(cè)細(xì)胞增殖情況,雙氯熒光素(DCFH-DA)探針檢測(cè)細(xì)胞活性氧(ROS)分子表達(dá),Western blotting法檢測(cè)細(xì)胞培養(yǎng)上清collagenⅠ、collagenⅢ和細(xì)胞中KCa3.1通道蛋白表達(dá)、PI3K信號(hào)通道蛋白磷酸化改變。結(jié)果:4、8和12月齡dTH小鼠CFs中ROS和KCa3.1通道蛋白表達(dá)水平與2月齡dTH小鼠比較均增加(P0.05或P0.01);MTT檢測(cè),應(yīng)用1×10~(-6)、1×10~(-5)、1×10~(-4)和1×10~(-3) mol·L~(-1) NAC干預(yù)后,dTH小鼠CFs增殖率分別為165.9%、138.72%、110.92%和109.82%,與dTH組比較,1×10~(-4)和1×10~(-3) mol·L~(-1) NAC對(duì)CFs增殖抑制作用明顯(P0.01)。與對(duì)照組比較,1×10~(-4) mol·L~(-1) NAC組小鼠CFs中Ⅰ、Ⅲ型膠原合成明顯減少(P0.01);Western blotting檢測(cè),與對(duì)照組比較,1×10~(-4) mol·L~(-1) NAC組小鼠CFs中KCa3.1通道蛋白表達(dá)水平下降(P0.01);dTH小鼠CFs中p-Akt/T-Akt表達(dá)水平較對(duì)照組增加(P0.01),而1×10~(-4) mol·L~(-1) NAC組小鼠CFs中p-Akt/T-Akt表達(dá)水平下降(P0.01)。結(jié)論:ROS清除劑NAC抑制dTH高血壓小鼠CFs中KCa3.1通道蛋白表達(dá),可能與其上調(diào)細(xì)胞中PI3K信號(hào)通道磷酸化水平有關(guān)。
[Abstract]:Aim: to investigate the relationship between the level of oxidative stress of cardiac fibroblasts (CFS) and the expression of medium conductance calcium activated potassium channel (KCA 3.1) protein during hypertension, and to elucidate the role and mechanism of KCa 3.1 in myocardial fibrosis. Methods: the primary cultured male AGT-REN double transgenic hypertensive mice were cultured with dTHH CFS, and the same strain of wild C57B6 mice as control. The CFs of the same strain of wild C57B6 mice were randomly divided into two groups: hypertension group (DTHs) and N- acetylcysteine group (N- acetylcysteine group) and NAC dry cells were given to different concentrations of CFS in the same strain of wild C57B6 mice. Cell proliferation was detected by 24 h. MTT assay. Dichlorofluorescein (DCFH-DAA) probe was used to detect the expression of reactive oxygen species (Ros) in cells. Western blotting assay was used to detect the phosphorylation of collagen 鈪，

本文編號(hào)：1993190