游离血红蛋白对体外培养成年大鼠嗅鞘细胞存活的影响  

Effect of free hemoglobin on viability of olfactory ensheathing ceils in vitro from adult rats

作　　者：薛海龙[1] 罗娜[1] 杨浩[1] 沈学锋[1] 鞠躬[1] 

机构地区：[1]第四军医大学神经生物学教研室,西安710032

出　　处：《中华神经医学杂志》2007年第1期9-12,共4页Chinese Journal of Neuromedicine

摘　　要：目的观察不同浓度游离血红蛋白（FHb）对体外培养大鼠嗅鞘细胞（OECs）存活的影响。方法采用成年大鼠（2．5月）嗅球培养OECs，培养第8天用S-100免疫组化鉴定纯度后，将培养的OECs根据加入的FHb浓度分为1，10，50，100μmol／L组及正常对照组，观察不同浓度FHb下OECs形态学变化；培养24h后采用MTT实验检测不同浓度FHb组细胞活力；并用碘化丙啶（PI）和Hocchst33342荧光双染色观察检测细胞死亡。结果大鼠OECs纯度为（72±6）％；MTT试验显示各组细胞活力随血红蛋白浓度的升高而降低；PI／Hoechst33342双荧光染色显示在对照组未见PI染色阳性细胞，各FHb组细胞PI／Hoechst33342比例随血红蛋白浓度增加而升高。结论FHb对OECs有毒性作用且随浓度的增加而增加。Objective To investigate the effect of free hemoglobin （FHb） on the viability of olfactory ensheathing cells （OECs） in vitro from adult rats. Metheds The OECs were dissociated fi＇om the olfactory bulbs of adult rats （2.5 months）. ARer 8 d of primary culture, the purity of OECs were evaluated according to the percentage of S-100 positive cells. The cultured OECs was divided into 5 groups each treated with different concentrations of FHb （1, 10, 50, 100 ixmol/L and control group）. The morphological change of OECs was examined under a phase contrast microscope. At 24 h, MTT assay was performed to assess the viability of OECs, and Propidium Iodide （PI）/Hocchst33342 double staining was employed to investigate the death rate of OECs. Results The purity of OECs reached （72±6）%. There was no obvious morphological changes of OECs in control group, and the viability of OECs was suppressed in FHb groups, and many died. ARer treatment with different concentrations of FHb for 24 h, the death rate of OECs rose with the increasing concentration of FHb. No cell death could be detected in the control group. PI / Hoechst33342 double staining showed the control group had no PI positive cells, but FHb groups had increasing PI/Hoechst33342 positive cells with the increasing concentration of FHb. Condusion FHb is toxic to OECs in a concentration dependent manner.

关 键 词：嗅鞘细胞 细胞存活 游离血红蛋白 

分 类 号：R686[医药卫生—骨科学]