色满卡林对培养乳鼠心肌细胞H/R损伤的保护作用研究

The effect of CRK on the damage induced by H/R in cardiac muscle cells

目的观察色满卡林(CRK)对培养的大鼠乳鼠心肌细胞缺氧复氧损伤的保护作用并探讨其作用机制。方法采用原代培养的SD大鼠乳鼠心肌细胞,随机分为5组:正常对照组:不施加任何处理因素正常培养;H/R组:缺氧2h,复氧30min;CRK各浓度组:分别加入终浓度为2.5、5、10μmol/L的CRK后均即刻缺氧2h、复氧30min。各组细胞采用乳酸脱氢酶(LDH)试剂盒测培养液中LDH的活性;AV-PI双标记后应用流式细胞仪检测心肌细胞的凋亡率;Western blot方法检测caspase-3蛋白的表达。结果 H/R组培养液中LDH活性、心肌细胞凋亡率较正常对照组明显升高,caspase-3蛋白表达水平上调。CRK各浓度组培养液中LDH活性和心肌细胞凋亡率较H/R组降低,caspase-3蛋白表达水平下调。结论 CRK对缺氧复氧致培养大鼠乳鼠心肌细胞的损伤具有保护作用,其机制可能为CRK维持细胞膜完整性与稳定性,下调caspase-3活性蛋白表达水平,减少心肌细胞凋亡率。

Objective To investigate the effects of CRK on the damage induced by H/R in cardiac muscle cells.Methods The cardiac muscle cells isolated from new born S D rats were cultured,then randomly divided into five groups:(i).Normal cultur es(control);(ii).H/R control cultures;(iii).H/R cultures pretreated with CRK(2.5、5、10 μmol/L).LDH leakage of cardiac muscle cells was detected with chrom atometry and flow cytometric analysis for identification and quantification of c ell apoptosis,and the excepression of caspase-3 was observed by western blot.Results After H/R cultures,the content of LDH in culture solution and the expres sion of caspase-3 in cardiac muscle cells were raised obviously,and cells apopt osis were increased(P<0.01),compared with normal group;CRK(2.5、5、10μmo l/ L) could improve cells` change induced by H/R,LDH leakage,the express of caspase-3 and apoptosis ratio were reduced in different degree.Conclusion CRK could pr otect cardiac muscle cells refrain from the damage induced by H/R effectively,t he mechanism may be that CRK could keep the integrity and stability of cell memb rance and keep the low level of caspase-3 to relieve cells apoptosis.