脑出血大鼠脑内嗅鞘细胞移植后生长相关蛋白-43及层粘连蛋白的表达变化

目的研究嗅鞘细胞移植对大鼠脑出血后脑内生长相关蛋白-43及层粘连蛋白表达的影响。方法取新生3d的Wistar乳鼠嗅球,差速贴壁法培养获得嗅鞘细胞。40只Wistar大鼠造模后(胶原酶Ⅶ注入尾状核)随机分为嗅鞘细胞移植组和脑出血模型组。嗅鞘细胞移植组(20只)术后3d,移植嗅鞘细胞;脑出血模型组(20只)注射等量的生理盐水。两组大鼠分别在术后1d、7d、14d及28d用Bederson方法进行评分。各时间点于每组随机取3只大鼠处死,取脑组织做成石蜡切片,免疫组化法观察生长相关蛋白-43及层粘连蛋白表达的变化。结果运动功能评分显示,移植组与对照组均出现运动功能恢复,移植组明显优于对照组(P<0.05);生长相关蛋白-43及层粘连蛋白免疫组化结果表明,除术前1d外,其它各时间点生长相关蛋白-43及层粘连蛋白阳性表达值,移植组较对照组均强,差异有统计学意义(P<0.05)。结论嗅鞘细胞移植可提高生长相关蛋白-43及层粘连蛋白的表达,改善脑出血后神经再生微环境,促进神经再生。

Changes of growth associated protein-43 and laminin expression after transplantation of olfactory ensheathing cells in rats of intracerebral hemorrhage

Objective To study the effect of growth associated protein-43(GAP-43)and laminin(LN)expression after olfactory ensheathing cells(OECs)transplant to rat with intracerebral hemorrhage(ICH).Methods OECs were obtained from the olfactory bulb of 3 days newborn Wistar rat,and the cells for transplantation were purified using differential time adherent method.Collagenase Ⅶ was injected into the caudate nucleus to establish the model of intracerebral hemorrhage.Forty Wistar rats of intracerebral cerebral hemorrhage models were established and divided into two groups,the first group(20 rats)was the transplanting group,the second group(20 rats)was the model group.3 days later,undifferentiated OECs were injected into the same side of cerebrum with hematoma in the first group,and the same amount of saline was injected in the second group for contrast.Two groups were both scored on 1d,Td,14d and 28d after operation by Bederson method,then three rots were killed randomly in all time points after operation in every group.Paraffin sections were prepared for immunohistochemical staining of GAP-43 and LN.Results Motor function score showed locomotion behavior deficit were improved in all group,and score of animals in transplanting group were significantly lower than that of control group in all periods except on 1d(P ＜ 0.05).The result of immunohistochemistry staining indicated that at all time points except 1d,the positive expressions of GAP-43 and LN in the transplanting group were higher than those of in the model group,and there were statistically significant differences between them.Conclusion OECs transplantation to treat intracerebral hemorrhage enhances the expression of GAP-43 and LN,improves the microenvironment of the nerve regeneration and promotes the nerve regeneration.