硝苯地平对过氧化氢引起的心肌纤维化的抑制作用及机制  

The anti-fibrotic effect and mechanism of nifedipine on H_2O_2-induced cardiac fibrosis

作　　者：贾岩岩[1] 陈少锐[1] 徐剑[1] 郭金蕾[1] 刘培庆[1] 蒋建敏[1] 

机构地区：[1]中山大学药学院药理与毒理学实验室

出　　处：《广东医学》2013年第9期1313-1317,共5页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:30670837);国家自然科学基金中加合作项目(编号:30811120434);广东省自然科学基金资助项目(编号:S2012040006327)

摘　　要：目的研究硝苯地平(nifedipine)对过氧化氢(H2O2)引起的心肌纤维化的影响及机制。方法原代培养大鼠心脏成纤维细胞(CFs),用100μmol/L H2O2刺激,蛋白免疫印迹法(Western blot)检测硝苯地平(10μmol/L)以及对照药维拉帕米(verapamil,10μmol/L)对H2O2诱导的结缔组织生长因子(CTGF)、纤维粘连蛋白(FN)的蛋白表达以及信号蛋白MAPK磷酸化的影响;利用fluo-4/AM钙荧光探针法测定硝苯地平对H2O2刺激引起CFs内钙离子浓度([Ca2+]i)变化的影响。结果 (1)硝苯地平能够显著抑制H2O2诱导的CTGF、FN蛋白表达上调以及细胞外信号调控蛋白激酶1/2(ERK1/2)、c-Jun N端激酶(JNK)磷酸化,而对p38MAPK磷酸化没有影响,维拉帕米对以上蛋白表达均无抑制作用;(2)硝苯地平对H2O2刺激引起的CFs中的[Ca2+]i增加没有影响;(3)ERK信号通路抑制剂PD98059(10μmol/L)可抑制H2O2诱导的CTGF、FN的蛋白表达上调。结论硝苯地平可抑制H2O2刺激引起的CTGF、FN蛋白表达上调,而对[Ca2+]i变化没有影响,其抗纤维化的作用不依赖于经典的钙离子阻断作用,可能与抑制ERK1/2磷酸化有关。Objective To study the effects of nifedipine on H2O2 - induced cardiac fibrosis and its mechanism. Methods Primal3, cultured rat cardiac fibroblasts （CFs） were stimulated with H202 （ 100 μM）. The effects of nifedipine on H2O2 -induced connective tissue growth factor （CTGF） , fibroneetin （FN） expression, as well as the phosphorylation of MAPK （mitogen activated protein kinases ） were identified by Western blot analysis. The selective fluorescent probe fluo - 4/AM was used to measure intracellular calcium concentration （ [ Ca2 ＋ ~ i ） changes. Results Nifedipine signifi- cantly depressed The H202 - induced up - regulation of CTGF and FN, as well as the phosphorylation of extraeellular sig- nal -regulate kinases 1/2 （ERK1/2） and c -Jun NH （2） -terminal kinase （JNK） , were significantly depressed by nifedipine ; no effect on H2O2 - induced [ Ca2 ＋ ] i increase in CFs was observed. The H2O2 - induced up - regulation of CTGF and FN was also depressed by ERK inhibitor PD98059, Conclusion Nifedipine inhibits H2O2 - induced cardiac fibrosis through interference with the ERK1/2 signaling pathway, but not via the classic calcium channel blocking.

关 键 词：硝苯地平 过氧化氢 心肌纤维化 信号通路 钙离子 

分 类 号：R544.105[医药卫生—心血管疾病]