α7烟碱乙酰胆碱受体激动剂可能通过抑制p38丝裂原活化蛋白激酶信号通路减少心肌纤维化  

α7 nicotinic acetylcholine receptor agonist reduced myocardial fibrosis possibly through inhibiting p38 mitogen activated protein kinase signaling pathway

作　　者：张燕[1] 符晓阳[2] 

机构地区：[1]河南省人民医院郑州大学人民医院心血管内科,郑州450003 [2]河南省人民医院郑州大学人民医院心血管外科,郑州450003

出　　处：《中华实验外科杂志》2017年第4期622-625,共4页Chinese Journal of Experimental Surgery

基　　金：2016年河南省科技攻关项目（162102310034）

摘　　要：目的 探讨α7烟碱乙酰胆碱受体（α7nAchR）激动剂在心肌纤维化中的作用及其机制。 方法 分离培养乳鼠心肌成纤维细胞，传代培养2～4代时，分为4组：空白对照组、模型组、α7nAchR激动剂组和α7nAchR阻断剂组。空白对照组不做任何处理，模型组仅加入10－6 mol/L的血管紧张素Ⅱ（Ang Ⅱ），α7nAchR激动剂组和α7nAchR阻断剂组分别加入5×10－6 mol/L的PNU-282987和10－6 mol/L的α7nAchR抑制剂甲基牛扁亭碱（MLA），60 min后加入10－6 mol/L的Ang Ⅱ。水溶性四唑盐WST-1检测不同处理组心肌成纤维细胞增殖，实时荧光定量聚合酶链反应（FQ-PCR）检测不同处理组心肌成纤维细胞中α7nAchR基因表达，Western blot法检测不同处理组心肌成纤维细胞中α7nAchR、Ⅰ型和Ⅲ型胶原蛋白、α-平滑肌肌动蛋白（α-SMA）、p38丝裂原活化蛋白激酶（p38MAPK）和磷酸化p38MAPK（p-p38MAPK）蛋白表达。结果 （1）α7nAchR激动剂组心肌成纤维细胞最终吸光度（0.50±0.13）显著低于α7nAchR阻断剂组和模型组（0.75±0.10、0.63±0.11，F＝10.567，P＝0.000），提示激动α7nAchR可抑制心肌成纤维细胞增殖。（2）α7nAchR激动剂组心肌成纤维细胞中α7nAchR mRNA和蛋白相对表达量（0.87±0.15、0.76±0.08）显著高于α7nAchR阻断剂组（0.45±0.09、0.40±0.14）、模型组（0.62±0.11、0.59±0.10）和空白对照组（0.32±0.13、0.30±0.07，F＝25.402，P＝0.000），提示α7nAchR激动剂可上调心肌成纤维细胞中α7nAchR基因表达。（3）α7nAchR激动剂组Ⅰ型和Ⅲ型胶原蛋白、α-SMA蛋白、p-p38MAPK蛋白相对表达量（0.53±0.09、0.50±0.12、0.38±0.08、0.27±0.09）均低于模型组（0.65±0.12、0.62±0.10、0.57±0.11、0.45±0.11）和α7nAchR阻断剂组（0.81±0.13、0.71±0.11、0.70±0.10、0.68±0.08），而高于空白对照组（0.41±0.08、0.35±0.06、0.19±0.07、0.16±0.07，F＝29.647、32.962、30.549、53.665，P＝0.000、Objective To investigate the role of α7 nicotinic acetylcholine receptor （α7nAchR） agonist in myocardial fibrosis and its possible mechanism. Methods The cardiac fibroblasts were isolated and cultured from neonatal rats. The cardiac fibroblasts were divided into the following four groups： blank control group （cardiac fibroblasts without any intervention）; model group [cardiac fibroblasts treated with 10-6 mol/L angiotensin Ⅱ （Ang Ⅱ） only]; α7nAchR agonist group （cardiac fibroblasts treated with 5×10-6 mol/L PNU-282987, and 1 h later treated with 10-6 mol/L Ang Ⅱ）; α7nAchR antagonist group （cardiac fibroblasts treated with 10-6 mol/L MLA, and 1 h later treated with 10-6 mol/L Ang Ⅱ）. The proliferation abilities of cardiac fibroblasts in different treatment groups were measured using WST-1 method. The expression levels of α7nAchR genes in cardiac fibroblasts in different treatment groups were examined by real-time fluorescent quantitative polymerase chain reaction （FQ-PCR）. The expression levels of α7nAchR, type Ⅰ and type Ⅲ collagen, α-smooth muscle actin （α-SMA）, p38 mitogen activated protein kinase （p38MAPK） and phosphorylated p38MAPK （p-p38MAPK） proteins in cardiac fibroblasts were detected by Western blotting. Results （1） The final absorbance in the α7nAchR agonist group （0.50±0.13） was significantly lower than α7nAchR antagonist group and model group （0.75±0.10, 0.63±0.11, F=10.567, P=0.000）. It suggested that activation of α7nAchR could inhibit the proliferation of cardiac fibroblasts. （2） The relative expression levels of α7nAchR mRNA and protein in α7nAchR agonist group （0.87±0.15, 0.76±0.08） were significantly higher than the α7nAchR antagonist group （0.45±0.09, 0.40±0.14）, model group （0.62±0.11, 0.59±0.10） and blank control group （0.32±0.13, 0.30±0.07, F=25.402, P=0.000）. It suggested that α7nAchR agonists can upregulate the expression of α7nAchR gene in cardiac fibroblasts. （3） The relati

关 键 词：心肌成纤维细胞 α7烟碱乙酰胆碱受体激动剂 纤维化 p38丝裂原活化蛋白激酶信号通路 

分 类 号：R54[医药卫生—心血管疾病]