促红细胞生成素联合粒细胞集落刺激因子对大鼠缺氧心肌细胞的保护效应研究  

The research of protective effects induced by erythropoietin in combination with granulocyte colony- stimulating factor on hypoxic myocardial cells in rat

作　　者：马建林[1] 郭文玲[2] 李宁[2] 

机构地区：[1]滨州市中心医院急诊科,251700 [2]滨州市中心医院产科,251700

出　　处：《国际免疫学杂志》2016年第2期143-145,共3页International Journal of Immunology

摘　　要：目的探讨促红细胞生成素（EPO）联合粒细胞集落刺激因子（G-CSF）对大鼠缺氧心肌细胞的保护机制。方法采用胰酶消化大鼠心肌细胞并传代，用150μmol／L浓度的CoCl2处理心肌细胞，模拟缺氧条件。采用流式细胞仪检测不同EPO＋G—CSF浓度下心肌细胞的凋亡和坏死水平。筛选出最适浓度后，将实验组分为对照组、缺氧组、缺氧＋EP0组、缺氧＋G—CSF组和缺氧＋EPO＋G-CSF组，Westernblot法检测乏氧诱导因子（HIF）-1α、P53、PARP蛋白的表达变化。结果心肌细胞在10U／mLEPO＋200ng／mLG—CSF培养下，心肌细胞凋亡和坏死水平最低，故认为是最适浓度（F=14．73、11．95，P〈0．05）。和对照组相比，HIF-1α、P53和PARP蛋白在各实验组中表达水平均升高，HIF-1α以缺氧组升高最显著（F=22．65，P〈0．05），P53以缺氧＋EPO＋G—CSF组升高最显著（F=25．18，P〈0．05）。除对照组外，PARP蛋白在实验组中呈现逐渐降低趋势（F=17．48，P〈0．05）。结论EPO-G-CSF能降低缺氧的心肌细胞凋亡、坏死和DNA损伤，其机制可能与HIF-1α、P53、PARP蛋白表达相关。Objective To explore the protective effects of erythropoietin （EPO） in combination with Granuloeyte eolony-stimulating factor（G-CSF） on hypoxic myocardial cells in rat. Methods Myocardial cells were digested with pancreatin and subcultured. Cells was treated with CoC1z under the condition of hypoxic. Flow cytometry was used to detect the changes of apoptosis and necrosis level in myocardial cells and the optimal con- centration was selected. The experimental groups were divided into control, hypoxia, hypoxia ＋ EPO, hypoxia ＋ G-CSF,hypoxia ＋ EPO ＋ G-CSF. Western blot was used to detect the protein expression of hypoxia-indueible factor（HIF）-1α,P53 and PARP. Results Myocardial cells showed the lowest level of apoptosis and necrosis under cultured with 10 U/EPO and 200 ng/LG-CSF（ F = 14. 73,11.95, P 〈 0. 05 ）. Compared with control, the protein expression of HIF-1α, P53 and PARP was increased in all experimental groups. The protein expression of HIF-1α and P53 was largely increased in hypoxia and hypoxia ＋ EPO ＋ G-CSF, respectively （ F = 22. 65, F = 25. 18 ,P 〈 O. 05 ）. The protein expression of PARP gradually decreased in experimental groups besides control （F = 17.48 ,P 〈 0. 05 ）. Conclusion EPO and G-CSF could decrease apoptosis, necrosis and DNA damage in hypoxic myocardial ceils. The protective effects was associated with the protein expression of HIF-1α, P53 and PARP.

关 键 词：促红细胞生成素 粒细胞集落刺激因子 缺氧 

分 类 号：R542.22[医药卫生—心血管疾病]