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作 者:魏子寒[1] 王颖[1] 杨国杰[1] 孙丽娜[1]
机构地区:[1]郑州大学第一附属医院老年心血管科,河南省郑州市450052
出 处:《中国循环杂志》2014年第9期738-742,共5页Chinese Circulation Journal
摘 要:目的:观察磷酸肌酸钠对血管紧张素Ⅱ(AngⅡ)诱导的乳鼠心肌成纤维细胞(CF)增殖和胶原合成的影响,并初步探索磷酸肌酸钠抗心肌纤维化的作用机制。方法:将20只Wistar乳鼠取出心脏,体外原代、传代培养CF。实验分4组(每组n=3),对照组:无血清的DMEM培养液培养CF;AngⅡ组:含AngⅡ1×10-6mol/L的无血清DMEM培养液;磷酸肌酸钠组:含磷酸肌酸钠10mmol/L的无血清DMEM培养液;AngⅡ+磷酸肌酸钠组:含磷酸肌酸钠10 mmol/L加AngⅡ1×10-6mol/L的无血清DMEM培养液。采用流式细胞术测定细胞周期分布,Van Gieson(VG)氏染色法测定胶原含量,免疫细胞化学法检测磷酸化细胞外信号调节激酶(pERK1/2)蛋白的表达水平。结果:与对照组相比,AngⅡ组CF的S期细胞百分率明显增加,G0/G1期、G2/M期细胞百分率降低,胶原含量增加,pERK1/2蛋白表达增高,差异均有统计学意义(P均<0.01)。与对照组比较,磷酸肌酸钠组CF细胞周期、胶原含量和pERK1/2蛋白表达,差异均无统计学意义(P均>0.05)。与对照组比较,AngⅡ+磷酸肌酸钠组pERK1/2蛋白表达升高,差异有统计学意义(P<0.01),CF细胞周期和胶原含量差异均无统计学意义(P均>0.05)。与AngⅡ组比较,AngⅡ+磷酸肌酸钠组G0/G1期、G2/M期细胞百分率升高,S期百分率降低,胶原含量减少,pERK1/2蛋白表达降低,差异均有统计学意义(P均<0.01)。结论:磷酸肌酸钠可部分抑制AngⅡ诱导的CF增殖和胶原合成增加,其机制可能与抑制ERK1/2过度激活有关。这提示磷酸肌酸钠可以明显改善AngⅡ诱导的心肌纤维化。Objective: To investigate the effect of phosphocreatine (PCr) on angiotensin Ⅱ (Ang Ⅱ) induced proliferation and collagen synthesis of cardiac ifbroblasts in neonatal rats with its mechanism. Methods: The cardiac ifbroblasts (CF) from neonatal rats were cultured in vitro and were divided into 4 groups.①Control group, the CF was cultured in non-serum DMEM,②Ang Ⅱ group, the CF was cultured with Ang Ⅱ at (1&#215;10-6) mol/L,③PCr treated group, the CF was cultured with PCr at 10 mmol/L, and④Ang Ⅱ+PCr group. The CF cell cycle percentage was detected by lfow cytometric assay, myocardial collagen content was observed by VG staining and protein expression of phosphorylated extracellular signal-regulated kinase (pERK1/2) was detected by immuneohistochemistry. Results: ① Compared with Control group, the CF in Ang Ⅱ group showed increased percentage of S phase and decreased percentage of G0/G1 and G2/M phases, increased collagen content and pERK1/2 protein expression, all P〈0.01.② The CF cell cycle, collagen content and pERK1/2 protein expression were similar between Control group and PCr treated group, all P〉0.05. ③ Compared with Control group, Ang Ⅱ + PCr group had elevated pERK1/2 protein expression, P〈0.01, while the CF cell cycle and collagen content were similar with Control group, P〉0.05.④Compared with Ang Ⅱ group, the CF in Ang Ⅱ + PCr group had increased percentage of G0/G1 and G2/M phases, decreased percentage of S phase, decreased collagen content and pERK1/2 protein expression, all P〈0.01. Conclusion: PCr may partially inhibit Ang Ⅱ induced CF proliferation and collagen synthesis which might be related to the inhibition of excessively activated ERK1/2. Therefore, PCr could improve Ang Ⅱ induced myocardial ifbrosis in neonatal rats.
关 键 词:磷酸肌酸钠 心肌 成纤维细胞 磷酸化细胞外信号调节激酶
分 类 号:R54[医药卫生—心血管疾病]
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