促红细胞生成素对缺氧乳鼠心肌细胞凋亡及坏死的影响

目的观察促红细胞生成素(EPO)对对缺氧心肌细胞的保护效应及量效关系。方法将当日生乳鼠,局部碘酒酒精消毒后胸骨正中开胸,取心尖前1/2心室肌进行细胞培养。培养4 d后选择细胞生长情况良好的培养瓶(>106个细胞/瓶)分为3组:A,对照组,正常培养;B,缺氧组,缺氧1h/复氧12 h;C,EPO组,缺氧/复氧加EPO干预。EPO组根据不同EPO干预浓度又分为C1,EPO干预浓度0.1 IU/ml;C2,EPO浓度1 IU/ml;C3,EPO干预浓度10 IU/ml。然后以荧光染色-流式细胞仪检测缺氧/复氧后心肌细胞的凋亡及坏死情况。结果缺氧组细胞的凋亡、坏死比率分别为(20.78±2.98)%和(50.10±0.29)%,明显高于EPO各组(P均小于0.01),也明显高于对照组[(2.3±1.09)%和(1.5±0.48)%,P<0.01]。不同浓度EPO浓度干预组间比较,C1组的存活率(40.18±7.49)%低于C2组、C3组(65.56±6.37)%,(62.56±11.32)%,均P<0.01,凋亡率(13.78±3.74)%和坏死率(42.51±5.49)%则明显高于后二者(P<0.05或P<0.01)。C2及C3间各指标差异无统计学意义。结论早期应用EPO可明显减少体外培养的乳鼠心室肌细胞缺氧/复氧后凋亡和坏死的发生。而且EPO在0.1 IU/ml至1 IU/ml浓度范围内其保护作用存在一定的量效关系。

Erythropoietin decreases apoptosis and death of cultured myocardial cell induced by hypoxia/reoxygenation.

Objective To study the effect of erythropoietin（EPO） on apoptosis and death in cultured cardiamyocytes after hypoxia and reoxygenation. Method Cultured cardiomyocytes from neonate rats were divided in three groups：control group（A）, hypoxia group （B, hypoxia for 2 h and reoKygenation for 12 h） and EPO group （C, hypoxia lh/reoxygenation 12h added with EPO）. According to different concentration of EPO, group C were further divided into three subgroups：C1, EPO 0. lIU/ml; C2,EPO lIU/ml; C3, EPO 10IU/ml. At the end of intervention, the cells were stained with fluorescent dyes and apoptosis and death were assayed by flow cytometry. Results 1. After hypoxia and reoxygenation, the percentage of apoptosis（ 20. 78± 2.98）% and necrosis （50.10 ± 0.29）% of hypoxia group were significantly higher than those of control group [ （ 8.41 ± 0.52 ） % and （ 18.56 ± 0.56 ） %, P 〈 0.01, respectively ] and EPO subgroups （ P 〈 0.01, respectively ）. Among EPO subgroups, death and necrosis rates （ 13.78 ± 3.74） % ; （42.51 ± 5.49） % of C1 were much higher than those of C2 and C3 （ P 〈 0.05 or P 〈 0.01 ）. There was no difference between group C2 and C3. Conclusion EPO may significantly decrease the apoptosis as well as necrosis of hypoxia/reoxygenation injured cardialmyocytes in a dose dependent manner within a range of concentration.