脑源性神经生长因子和神经干细胞联合移植对大脑中动脉阻塞大鼠神经功能恢复的作用

目的 将脑源性神经生长因子(BDNF)和神经干细胞(NSCs)单独及联合移植应用于大脑中动脉阻塞(MCAo)模型大鼠,观察BDNF和NSCs移植对大鼠缺血性脑卒中神经功能恢复的作用及BDNF对内源性和外源性NSCs增殖、迁移及分化的影响.方法 体外分离、培养新生大鼠海马NSCs,BrdU标记.实验动物随机分为A组(MCAo组);B组(MCAo+BDNF组);C组(MCAo+NSCs组);D组(MCAo+BDNF+NSCs组),每组16只,移植后进行神经功能损害评分(NSS),用免疫组织化学行BrdU、nestin、BrdU/NSE检测,分析结果.结果 移植后的2、4周神经功能评分分别为:A组5.3±0.5、5.3±0.5;B组4.0±0.8、3.8±0.5;C组3.5±0.6、3.5±0.6;D组2.0 ±0.8、1.8±1.0,D组显著好于其他3组(P<0.05),B组与c组显著好于A组(P<0.05).nes.tin阳性细胞数:A组1.24±1.13,B组2.59±1.44(P<0.05),BrdU阳性细胞数:A组0.52±0.68,B组1.65±1.10(P<0.05).BrdU阳性细胞数:C组6.08±1.52,D组10.26±1.96(P<0.05),BrdU/NSE双阳性细胞数:C组1.74±1.04,D组3.58±1.20(P<0.05).结论 BDNF和NSCs移植单独及联合应用对MCAo大鼠的神经功能恢复均有作用,两者联合具有协同作用.BDNF对内源性NSCs的激活、增殖有促进作用,对外源性NSCs的增殖、迁移及分化有促进作用.

Effect of BDNF and NSCs transplantation on neurological function recovery after MCAo in rats

Objective The brain-derived neurotrophic factor (BDNF) and neural stem cells (NSCs) were respectively or jointly implanted into transient middle cerebral artery occlusion (MCAo) rats to study the effects of BDNF and NSCs transplantation on neurological function recovery in rats with ischemic stroke. Methods The hippocampal NSCs were isolated from neonate rats, cultured and labeled with BrdU. The model rats were randomly divided into 4 groups: group A (model group) ,group B (model and BDNF) ,group C (model and NSCs transplantation) and group D (model and BDNF, NSCs transplantation) (n = 16 in each group). The neurological deficits were evaluated with Neurological Severity Score (NSS) after transplantation. The brain was sectioned for immunohistochemical and immunofluoreseent staining. The statistical result were analyzed by a computer. Results The NSS at 2nd and 4th week after transplantation were 5.3±0.5 and 5.3 ±0.5 in group A,4.0±0.8 and 3.8±0.5 in group B,3.5 ±0.6 and 3.5±0.6 in group C, and 2.0±0.8 and 1.8±1.0 in group D, respectively. The NSS in group D were significantly lower than other other three groups (P < 0.05 ) and those in groups B and C were obviously lower than in group A ( P < 0.05 ). The number of BrdU-labeled and nestin-positive cells was (0.52 ± 0.68) and ( 1.24 ±1.13) in group A and ( 1.65±1.10) and (2.59 ±1.44) in group B, respectively (P < 0.05 ). The number of BrdU-labeled and BrdU/NSE double-positive cells was (6.08 ±1.52) and (1.74±1.04) in group C and ( 10.26 ±1.96) and (3.58 ± 1.20) in group D, respectively (P < 0.05). Conclusion The BDNF and NSCs transplantation respectively and jointly can improve the neurological function of rats after MCAo. The combined transplantation of BDNF and NSCs had a synergic effect. The BDNF can promote the endogenous NSCs activation and prolifeation and also improve the proliferation,migration and differentiation of the exogenous NSCs.