模拟高住低练对大鼠外周血内皮祖细胞增殖能力的影响

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目的:观察体外原代培养模拟高住低练后大鼠外周血内皮祖细胞的生长情况,探讨低氧与运动复合刺激对其增殖能力的影响.方法:选用健康雄性SD大鼠24只,随机分为4组:常氧居住组,低氧居住组,常氧训练组和高住低练组.低氧居住组采用递减氧浓度刺激,常氧训练组采用递增负荷跑台训练,高住低练组采用递减氧浓度刺激和递增负荷跑台训练,每周训练6天,为期8周.方案结束后12h提取大鼠外周血,采用密度梯度离心法分离外周血得到单个核细胞,以条件培养基对其体外培养诱导分化获取内皮祖细胞,通过双荧光染色法观察细胞对乙酰化低密度脂蛋白的摄取和与荆豆凝集素1的结合以及形态学特征鉴定内皮祖细胞.倒置显微镜下计数贴壁细胞数量和集落数目以反映细胞增殖情况.结果:与常氧居住组相比,其余三组贴壁细胞数量和集落数目显著增加(P<0.01),高住低练组贴壁细胞数量显著高于常氧训练组和低氧居住组(P0.05).结论:低氧和运动都能促进内皮祖细胞增殖,低氧和运动复合刺激效果更明显.“,”Objective We investigated if the proliferative capacity of endothelial progenitor cells was affected by hypoxia and exercise. Methods Twenty four male Sprague-Dawley rats were randomly and averagely divided into 4 groups: (1) living at low altitude (LL), (2) living and training at low altitude (LLTL), (3) living at high altitude (LH), and (4) living at high altitude and training at low altitude (LHTL). Eight-week incremental treadmill exercise and hypoxic simulation were used to establish LHTL animal model. Mononuclear cells from peripheral blood were obtained by density gradient centrifugation 12 hours by the end of 8-week experiment. The cells were suspended in conditioned medium 199 for culturing in vitro. Their phenotypes were confirmed by uptake of acetylated LDL and binding of fluoresce in isothiocyanate (FITC)-labeled Ulex europaeus agglutinin 1 (UEA-1) lectin. Inverted microscopic observation was used to identify the morphological changes in endothelial progenitor cells and measure the cell count. Results Adherent cells and early CFUs in groups LLTL, LH and LHTL increased more obviously than in group LL(P0.05). Conclusion Proliferative capacity of endothelial progenitor cells can be promoted by both hypoxic stimulation and exercise, and the promotion is more significant if combination of hypoxia and exercise was employed simultaneously.
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