α7nAChR激动剂对血管紧张素Ⅱ诱导的心肌成纤维细胞增殖和胶原合成的影响及其分子机制  

Effect of α7 Nicotinic Acetylcholine Receptor Agonist on AngiotensinⅡ-induced Cardiac Fibroblasts Proliferation and Collagen Synthesis

作　　者：符宇[1] 谈智[2,3] 吴涛[1] 耿登峰[1] 

机构地区：[1]中山大学孙逸仙纪念医院心内科,广东广州510120 [2]中山大学中山医学院生理教研室,广东广州510080 [3]中山医学院高血压研究所,广东广州510080

出　　处：《中山大学学报（医学科学版）》2016年第1期23-28,共6页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81270377);广东省自然科学基金(2014A030313092;2014A030313062)

摘　　要：【目的】探讨α7烟碱乙酰胆碱受体（α7 n Ach R）激动剂对血管紧张素Ⅱ（AngⅡ）诱导的心肌成纤维细胞增殖和胶原合成的影响及其分子机制。【方法】培养出生2~3d大鼠心肌成纤维细胞（CF）,取传代2~4代细胞,分为1空白对照组（control）;2模型组（AngⅡ10-7 mol/L）;3ACh预处理组：ACh＋AngⅡ组（ACh 10-4 mol/L＋AngⅡ10-7 mol/L）;4α7n ACh R激动剂组：α7n ACh R激动剂＋AngⅡ组（PNU-282987 5×10-6 mol/L＋AngⅡ10-7 mol/L）;5α7n ACh R拮抗剂组：ACh＋甲基牛扁碱柠檬酸盐（MLA）＋AngⅡ组（ACh 10-4 mol/L＋AngⅡ10-7 mol/L＋MLA 1×10-6 mol/L）。干预24 h后,应用CCK-8试剂盒检测CF的增殖能力,western blot检测胶原蛋白Ι、α-SMA、α7烟碱乙酰胆碱受体（α7n ACh R）、核中p65蛋白表达;Real-time PCR检测α7n ACh R的m RNA表达。【结果】AngⅡ干预24h,CF增殖显著提高,胶原蛋白Ι、α-SMA、核中p65蛋白的表达明显升高,α7n ACh R的m RNA和蛋白水平均表达降低;Ach、α7n ACh R激动剂预处理后抑制CF增殖,胶原蛋白Ι、α-SMA和核中p65蛋白的表达降低,α7n ACh R的m RNA和蛋白水平均表达升高;Ach＋MLA预处理,CF增殖再次提高,胶原蛋白Ι、纤维化因子α-SMA、核中p65蛋白的表达再次明显升高,α7n ACh R的m RNA和蛋白水平均表达降低。【结论】α7n ACh R激动剂抑制AngⅡ诱导的心肌成纤维细胞的增殖和胶原合成,其机制可能与NF-κB信号通路有关。【Objective】 To investigate the effect of an α7 nicotinic acetylcholine receptor（α7 n Ach R） agonist on cardiac fibroblast（CF） proliferation and collagen synthesis induced by angiotensin II（Ang II） and to determine its molecular mechanism.【Methods】 CF were cultured from hearts of Sprague-Dawley rats. The CF were divided into the following five groups ： 1 control group; 2 model group（Ang II 10-7mol / L）; 3AChpreconditioning group： ACh ＋ Ang II group（ACh 10-4mol / L ＋ Ang II10-7mol /L）; 4 α7n ACh R agonist group： α7n ACh R agonist ＋ Ang II group（PNU-282987 5×10-6mol / L ＋ Ang II10-7 mol / L）; 5 α7n ACh R antagonist group： ACh ＋ Methyllycaconitine citrate（MLA） ＋ Ang II group（ACh 10-4mol / L ＋ Ang II10-7 mol / L ＋ MLA 10-6mol / L）.The CCK-8 assay was used to test the proliferation of CF. Protein expression of collagen I, α-smooth muscle actin（α-SMA）,α7n ACh R, and p65 were analyzed by western blotting. The m RNA expression of α7n ACh R was measured by real-time PCR.【Results】 Incubation with Ang II induced significant proliferation of CF. Expressions of collagen I, α-SMA, and p65 were significantly increased, while the α7n ACh R protein and m RNA levels were significantly decreased. PNU-282987 significantlyinhibited the Ang II-induced CF proliferation; deregulated expressions of collagen I, α-SMA, and p65; and up-regulated α7n ACh R protein and m RNA levels. Preconditioning with ACh ＋ MLA restored the proliferation of CF and the expressions of collagen I, fibrosis factor α-SMA, and p65, while the α7n ACh R protein and m RNA levels decreased. 【Conclusion】 Anα7n ACh R agonist can inhibit Ang II-induced CF proliferation and collagen synthesis, in part by inhibiting the NF-κB signaling pathways.

关 键 词：心肌成纤维细胞 血管紧张素Ⅱ 胆碱能抗炎通路 α7烟碱乙酰胆碱受体 

分 类 号：R54[医药卫生—心血管疾病]