促红细胞生成素联合粒细胞集落刺激因子对乳鼠缺氧心肌细胞的保护作用.docVIP

促红细胞生成素联合粒细胞集落刺激因子对乳鼠缺氧心肌细胞的保护作用 付振虹，董蔚，盖鲁粤 （北京解放军总医院心血管内科心血管内科erythropoietin，EPO）联合粒细胞集落刺激因子（granulocyte colony stimulating factor，G-CSF）对缺氧心肌细胞的保护作用。方法 分离乳鼠心肌细胞。采用95% N2+5% CO2气体充分置换的1%的胎牛血清DMEM培养液培养24 h，建立缺氧心肌细胞模型。研究不同浓度的EPO和G-CSF对缺氧心肌细胞的保护作用。观察在最佳作用浓度时，EPO、G-CSF和联合用药对缺氧心肌细胞的保护作用。采用流式细胞仪检测各组心肌细胞的存活率、凋亡率和坏死率。结果 缺氧心肌细胞的死亡率（26.73%）和其中凋亡细胞与总死亡细胞的比例（70.05%）明显高于正常心肌细胞（5.63%、37.83%）。EPO 1 U/ml浓度开始发挥对缺氧心肌细胞的保护作用，5、25和125 U/ml的保护作用相似；G-CSF 6 ng/ml开始发挥对缺氧心肌细胞的保护作用，150 ng/ml保护作用最强，30、750 ng/ml保护作用相似。EPO（5 U/ml）、G-CSF（30 ng/ml）和联合用药[ EPO（5 U/ml）＋G-CSF（30 ng/ml）]明显减少缺氧心肌细胞的凋亡率 [（6.01±2.10）±0.16)、(4.57±0.99)，与缺氧心肌细胞（18.73±1.08）比较，P 0.01]和总死亡率[(9.54±3.51)、（11.6±0.65、（8.15±3.16）±1.30）P 0.01）；EPO组与G-CSF组保护作用相近（P 0.05）；联合给药组保护作用明显强于单独EPO与G-CSF给药组（P 0.01）。结论 EPO和G-CSF对缺氧心肌细胞具有保护作用，5 U/ml的EPO和30 ng/ml的G-CSF是减轻缺氧心肌细胞损伤的最佳剂量。者联合给药，保护效果优于单独用药。 关键词：缺氧，心肌细胞，促红细胞生成素，粒细胞集落刺激因子，细胞凋亡 Effect of Erythropoietin in Combination with Granulocyte- Colony Stimulating Factor on Hypoxia cardiomyocytes FU Zhen-hong, DONG Wei, GAI Lu-yue（PLA General hospital，Beijing 100853，China） Abstract Objective To evaluate the potential protective effects of EPO in combination with G-CSF on hypoxia cardiomyocytes. Methods Left ventricular cardiomyocytes were isolated from neonate rat, the air in the cultural medium was changed by 95%N2 and 5%CO2 for hypoxia cardiomyocytes. The protective effects of EPO and G-CSF in different concentrations were evaluated. Under the optimized concentration, the protective effects of EPO in combination with G-CSF were investigated. The survival, apoptotic and necrotic rates of cardiomyocytes were assessed by flow cytometry. Results The mortality and the rate of apoptotic cell to total necrotic cell were higher in hypoxia cardiomyocytes than normal cells significantly (26.73% vs. 5.63%，70.05% vs. 37.83%，P 0.01). At the concentration of 1U/ml of EPO and 6ng/ml of G-CSF, the protective effects occurred, 5U/ml, 25U/ml, and 125U/ml of EPO were similar, 150ng/ml of G-CSF was the most