CAJ | 학술논문

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10 mmol/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。
목적：관찰린산기산납대혈관긴장소Ⅱ（AngⅡ）유도적유서심기성섬유세포(CF)증식화효원합성적영향，병초보탐색린산기산납항심기섬유화적작용궤제。 　　방법：장20지Wistar유서취출심장，체외원대、전대배양CF。실험분4조（매조n=3），대조조：무혈청적DMEM배양액배양CF；AngⅡ조：함AngⅡ1×10-6mol/L적무혈청DMEM배양액；린산기산납조：함린산기산납10 mmol/L적무혈청DMEM배양액；AngⅡ+린산기산납조：함린산기산납10 mmol/L 가AngⅡ1×10-6mol/L적무혈청DMEM배양액。채용류식세포술측정세포주기분포，Van Gieson（VG）씨염색법측정효원함량，면역세포화학법검측린산화세포외신호조절격매（pERK1/2）단백적표체수평。 　　결과：여대조조상비，AngⅡ조CF적S기세포백분솔명현증가，G0/G1기、G2/M기세포백분솔강저，효원함량증가， pERK1/2단백표체증고，차이균유통계학의의（P균<0.01)。여대조조비교，린산기산납조CF세포주기、효원함량화pERK1/2단백표체，차이균무통계학의의（P균>0.05)。여대조조비교，AngⅡ+린산기산납조pERK1/2단백표체승고，차이유통계학의의（P<0.01)，CF세포주기화효원함량차이균무통계학의의（P균>0.05)。여AngⅡ조비교，AngⅡ+린산기산납조G0/G1기、G2/M기세포백분솔승고，S기백분솔강저，효원함량감소，pERK1/2단백표체강저，차이균유통계학의의（P균<0.01)。 　　결론：린산기산납가부분억제AngⅡ유도적CF증식화효원합성증가，기궤제가능여억제ERK1/2과도격활유관。저제시린산기산납가이명현개선AngⅡ유도적심기섬유화。
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 (1×10-6) 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.