N-乙酰半胱氨酸通路对AGT-REN双转基因高血压小鼠心肌成纤维细胞中电导钙激活钾离子通道蛋白表达的影响及其意义

发布时间：2018-06-07 23:05

  本文选题：心肌纤维化 + 活性氧　； 参考：《吉林大学学报(医学版)》2017年05期

【摘要】：目的:探讨高血压过程中心肌成纤维细胞(CFs)氧化应激水平与中电导钙激活钾离子通道(KCa3.1)蛋白表达的关系,阐明KCa3.1在心肌纤维化中的作用及机制。方法:原代培养雄性AGT-REN双转基因高血压小鼠(dTH)CFs,另培养同品系野生C57B6小鼠CFs作为对照。dTH小鼠CFs随机分为高血压组(dTH)和N-乙酰半胱氨酸组(NAC),dTH小鼠CFs给予不同浓度NAC干预24h。MTT法检测细胞增殖情况,双氯荧光素(DCFH-DA)探针检测细胞活性氧(ROS)分子表达,Western blotting法检测细胞培养上清collagenⅠ、collagenⅢ和细胞中KCa3.1通道蛋白表达、PI3K信号通道蛋白磷酸化改变。结果:4、8和12月龄dTH小鼠CFs中ROS和KCa3.1通道蛋白表达水平与2月龄dTH小鼠比较均增加(P0.05或P0.01);MTT检测,应用1×10~(-6)、1×10~(-5)、1×10~(-4)和1×10~(-3) mol·L~(-1) NAC干预后,dTH小鼠CFs增殖率分别为165.9%、138.72%、110.92%和109.82%,与dTH组比较,1×10~(-4)和1×10~(-3) mol·L~(-1) NAC对CFs增殖抑制作用明显(P0.01)。与对照组比较,1×10~(-4) mol·L~(-1) NAC组小鼠CFs中Ⅰ、Ⅲ型胶原合成明显减少(P0.01);Western blotting检测,与对照组比较,1×10~(-4) mol·L~(-1) NAC组小鼠CFs中KCa3.1通道蛋白表达水平下降(P0.01);dTH小鼠CFs中p-Akt/T-Akt表达水平较对照组增加(P0.01),而1×10~(-4) mol·L~(-1) NAC组小鼠CFs中p-Akt/T-Akt表达水平下降(P0.01)。结论:ROS清除剂NAC抑制dTH高血压小鼠CFs中KCa3.1通道蛋白表达,可能与其上调细胞中PI3K信号通道磷酸化水平有关。
[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，

本文编号：1993190