NGF联合神经干细胞移植对大鼠脑出血的治疗作用

Effect of transplantation of neural stem cells combined with nerve growth factor on intracerebral hemorrhage in rats

目的观察神经生长因子(Nerve growth factor,NGF)联合神经干细胞(Neural stem cells,NSCs)移植对脑出血后大鼠神经功能恢复的作用及NGF对移植NSCs的影响。方法 120只SD大鼠随机分为NSCs+NGF组、NSCs组、NGF组、模型对照组、空白对照组。体外分离、培养新生大鼠神经干细胞并进行染色,采用注射Ⅳ型胶原酶建立脑出血模型,造模后第3d行神经干细胞移植及NGF注射。移植后于相应时间点对大鼠进行神经行为学评分,后处死大鼠,测量血肿体积,观察各组大鼠脑组织HE染色及免疫组织化学染色。结果细胞移植后14d,大鼠神经功能缺损明显改善(P<0.05),NSCs+NGF组在移植28d时神经功能评分显著低于其他组(P<0.05)。细胞移植后14d,NSCs+NGF组大鼠血肿体积显著小于其他组(P<0.05)。细胞移植后7d,免疫组织化学染色显示NSCs+NGF组Brd U阳性细胞数显著高于NSCs组。结论 NSCs移植和NGF单独及联合应用对脑出血大鼠的神经功能恢复均有积极作用,二者联合具有协同作用。NGF对移植NSCs的增殖具有促进作用。

Objective To observe the effect of neural stem cells（NSCs） transplantation and nerve growth factor（NGF） on neurological function recovery in rats with intracerebral hemorrhage（ICH）. Methods One hundred and twenty male SD rats were employed and randomly divided into 5 groups（n=24）, namely NSCs＋NGF group, NSCs group, NGF group, model control group and blank control group. Cultured NSCs isolated from neonatal rats were labeled with BrdU. The IV collagenase was injected into basal nuclei to establish ICH model. Each rat was subjected to Score of Longa 5-grade standard to evaluate neurological function. The volume of hematoma of each rat was measured. Transplanted NSCs were detected by immunohistochemistry with double-labeling of BrdU and GFAP/NeuN. Results The neurological deficits of rats with ICH improved significantly at days 14 （P 〈 0.05）. NSCs ＋NGF group showed better functional performance 28 days after compared with other groups （P 〈 0.05） and the volume of hematoma were also significantly smaller compared with other groups after 14 days （P 〈 0.05）. BrdU ＋ ceils migrated selectively to the perihematomal areas and differentiated into neurons and astrocytes. Conclusion Both NSCs transplantation and NGF treatment can obviously improve the neurological function of rats with ICH, but their combined use can result in synergistic effect. NGF has stimulatory effect on the proliferation and activation of transplanted NSCs.