促红细胞生成素对过氧化氢损伤后大鼠骨骼肌卫星细胞凋亡和钙离子浓度的影响

Effect of erythropoietin on apoptosis and calcium ion concentration of skeletal muscle satellite cells injured by hydrogen peroxide in rats

目的探讨过氧化氢（H2O2）对大鼠骨骼肌卫星细胞（skeletalmusclesatellitecell，sMSC）凋亡和细胞内钙离子浓度的影响及促红细胞生成素（EPO）的保护作用。方法取体外培养的SMSC，随机分为对照组、Hz0：组和EPO干预组及EPO＋H2O2组，采用流式细胞仪检测细胞凋亡率和钙离子浓度的平均荧光强度，Hoechst33258染色后荧光显微镜观察凋亡细胞的形态学变化。结果H：02组凋亡发生率最高，为（22．13±1．79）％，经10、20、40u／m1的EPO预处理后凋亡率分别为（16．47±2．53）％、（4．97±0．55）％、（2．93±0．47）％，对照组凋亡率为（1．93±0．57）％；检测细胞内钙离子荧光强度结果显示，经10、20、40u／ml浓度的EPO预处理后分别为（12．67±0．32）、（27．90±0．66）、（44．53±0．93），与H2O2组（9．70±0．09）及对照组（51．37±0．64）比较，差异有统计学意义（P〈0．05）；H2O2组和低剂量EPO干预组（10U／ml、20U／ml），Hoeehst33258染色呈明显的凋亡征象，而高剂量EPO干预组（40u／ml）和对照组则较少见凋亡细胞。结论EPO可抑制H2O2诱导的细胞凋亡和减少钙离子的释放，且呈剂量相关性。

Objective To investigate the effect of hydrogen peroxide （H2O2） on apoptosis and calcium ion concentration of skeletal muscle satellite cells （SMSCs） in rats, and to explore the protective effect of erythropoietin （EPO）. Methods The cultured SMSCs were divided into five groups： control group, H2O2 group, 10, 20 and 40 U/ml EPO intervention groups. Apoptosis rates and calcium ion concentration of SMSCs were analyzed by flow cytometry, and the morphology of apoptotic cells was observed by Hoechst33258 staining. Results The apoptosis rates showed significant differences （all P〈0.05） among （1.93±0.57）% in control group, （22.13± 1.79）% in H202 group, （16.47±2.53） %, （4.97±0.55）% and （2.93±0.47）% in 10, 20 and 40 U/ml EPO intervention groups, respectively. And calcium ion concentrations in SMSCs were 12.67 ± 0.32, 27.90±0.06 and 44.53±0.93 in 10, 20 and 40 U/ml EPO intervention groups, respectively. There was significant difference in calcium ion concentration between H2 O2 group and control group （9.70± 0.09 vs. 51.37±0.64, P〈0.05）. Morphology of apoptosis was observed by Hoechst33258 dye stains in 10, 20 U/ml EPO intervention group and H2O2 group, while there were less apoptotic bodies in 40 U/rnl EPO intervention group and control group. Conclusions EPO might have protective effects on SMSCs injured by H2O2 through inhibiting apoptosis and calcium ion releasing from SMSCs.