嗅球和嗅黏膜来源的嗅鞘细胞对脊髓损伤小鼠的治疗效果比较  

Comparison of therapeutic effects of olfactory ensheathing cells derived from olfactory mucosa or olfactory bulb on spinal cord injury mouse models

作　　者：王立斌[1] 杨萍[1] 梁雪云[1] 马丽君[1] 魏军[1] 

机构地区：[1]宁夏医科大学总医院宁夏人类干细胞研究所,检验学院生育力保持教育部重点实验室,宁夏银川750004

出　　处：《细胞与分子免疫学杂志》2014年第4期379-383,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：宁夏自治区科技支撑计划项目资助(2013)

摘　　要：目的体外分离、培养不同来源的嗅鞘细胞,并鉴定其生物学特性,比较不同来源的嗅鞘细胞生物活性,评价其对脊髓损伤模型小鼠的疗效的差异。方法差速贴壁法分别培养嗅球和嗅黏膜来源的嗅鞘细胞,免疫荧光染色检测该细胞特异性蛋白S100、P75的表达,并对二者生长曲线及在不同代次、不同浓度梯度条件下神经营养因子分泌情况进行检测,实时定量PCR(qRT-PCR)鉴定二者脑源性神经营养因子(BDNF)、神经生长因子(NGF)、神经营养因子3(NT-3)、神经营养因子4(NT-4)、轴突膜蛋白生长相关蛋白43(GAP-43)、微管相关蛋白(MAP-2)基因表达,并分别将两种细胞移植入脊髓横断小鼠模型中,以小鼠神经功能评分及脊髓内移植细胞分布情况进行评估。结果所获得不同来源的嗅鞘细胞呈双极、三极样形态生长,且S100、P75蛋白表达均为阳性,嗅黏膜来源细胞不表达MAP-2,高表达GAP-43,嗅球来源的细胞低表达MAP-2和GAP-43,嗅球来源细胞生长活性大于嗅黏膜来源的细胞,其中嗅黏膜细胞来源分泌营养因子高于嗅球来源的细胞,小鼠神经功能评分显示嗅黏膜来源细胞治疗脊髓损伤模型效果明显高于嗅球来源的嗅鞘细胞。结论两种来源的嗅鞘细胞存在生物学差异,嗅黏膜来源细胞治疗脊髓损伤效果高于嗅球来源的嗅鞘细胞,可以作为临床应用的种子细胞。Objective To isolate and culture olfactory ensheathing cells from different origins, compare their different biological characteristics, and evaluate their therapeutic effect on spinal cord injury mouse models. Methods The olfactory ensheathing cells from olfactory mucosa or olfactory bulb were isolated and cultured by differential adhesion method. The expressions of S100 and P75 proteins were examined by immunofluorescence staining; their growth curves were drawn by Ml-r colorimetric assay; the secretion of neurotrophic factors, brain-derived neurotrophic factor （ BDNF）, nerve growth factor （NGF）, and neurotrophin-3 （NT-3） was measured by ELISA; the gene expressions of BDNF, NGF, NT-3, neurotrophin-4 （NT-4）, growth-associated protein 43 （GAP-43）, and microtubule-associated protein （MAP-2） were quantified by real-time PCR; the therapeutic effect on spinal cord injury mouse models was evaluated by Basso, Beattie and Bresnahan （BBB） locomotor rating scale, which had been carried out daily for 8 weeks after the olfactory ensheathing cells of the two different origins were respectively grafted to the mouse models. Results The two types of olfactory ensheathing cells showed bipolar or tripolar shape; both of them were S]00 and P75 protein positive; both of them expressing the gene of BDNF, NGF, NT-3, and NT-4; the olfactory bulb-derived cells did not express MAP-2, but it highly expressed GAP-43 gene; the olfactory mucosa- derived cells displayed a low expression of MAP-2 and GAP-43; the growth speed of olfactory bulb-derived cells was faster than that of the olfactory mucosa-derived cells. Both of them could secrete BDNF, NGF, and NT-3, but the neurotrophic factor levels secreted in the olfactory mucosa-derived cells were higher. The daily neurological BBB scoring showed that the therapeutic effect of olfactory mucosa-derived cells on spinal cord injury mouse models was better than that of the olfactory bulb-derived cells. Conclusion There exist biological differences between the olfactory

关 键 词：嗅鞘细胞 嗅球 嗅黏膜 脊髓损伤 

分 类 号：R392.31[医药卫生—免疫学] R392.33[医药卫生—基础医学]