低氧对大鼠骨髓基质细胞分泌神经生长因子的影响  

Expression of neurotrophic growth factor in rat bone marrow stromal cells cultured in hypoxia

作　　者：于红[1] 席焕久[2] 邓成国[1] 杨虹[1] 

机构地区：[1]郧阳医学院组织学与胚胎学教研室,湖北十堰442000 [2]辽宁医学院人类学研究所,辽宁锦州121001

出　　处：《解剖学杂志》2009年第3期316-318,共3页Chinese Journal of Anatomy

基　　金：辽宁省教育厅高等学校科学研究项目（05L129）

摘　　要：目的：研究大鼠骨髓基质细胞（BMSCs）在低氧培养条件下神经生长因子（NGF）的表达。方法：采用全骨髓贴壁培养法体外分离、纯化大鼠的BMSCs。置于常氧和低氧（3％）条件下培养，用cell counting kit（CCK-8）N定细胞增殖情况；台盼蓝染色检测细胞的活力；提取2种培养条件5个时间点的细胞蛋白，免疫印迹法检测细胞蛋白中NGF的含量；收集2种培养条件下5个时间点的细胞上清液，ELISA法检测细胞上清液中NGF的含量。结果：在3％低氧条件下培养BMSCs，其细胞活力较好，细胞增殖加快；免疫印迹法检测结果表明，与常氧条件相比，随培养时间的延长，低氧条件下NGF的表达明显增加，其24h分泌达高峰；ELISA检测结果表明，随培养时间的延长，常氧下细胞上清液中NGF的浓度逐渐增加，低氧12h时NGF的浓度显著增高，明显高于常氧培养组。结论：BMSCs在3％低氧条件下培养细胞生长迅速，细胞活力较好；3％的低氧条件可增强BMSCs分泌NGF的能力。Objective： To investigate expression of neurotrophic growth factor （NGF） in bone marrow stromal cells （BMSCs） of rat cultured in hypoxia. Methods： BMSCs from whole bone marrow of rat were cultured under normoxia and hypoxia. Cell count kit （CCK-8） was used to examine cell proliferation. Trypan blue dying was used to detect viability. Western blot was used to detect concentration of NGF in cells. NGF collected in cellular supernatant liquid was detected by ELISA. Resuits： The expression of NGF detected by Western blot was gradually enhanced in normoxia, but the tendency was not significant, while in hypoxia it obviously increased and reached peak at 24th hour; in normoxia, the concentration of NGF in the cellular supernatant liquid detected by ELISA gradually increased, while the concentration of NGF obviously increased at 12^th hour and was significantly higher than that innormoxia. Conclusion： 3% oxygen has an advantage to BMSCs with satis-factory proliferation and viability, and can enhance its secretion of NGF.

关 键 词：低氧 骨髓基质细胞 神经生长因子 大鼠 

分 类 号：R285.5[医药卫生—中药学] R318[医药卫生—中医学]