人骨髓间充质干细胞脑内移植治疗锰中毒模型的可行性

摘要

背景:研究证明骨髓间充质干细胞可以治疗中枢神经系统疾病,具有进行自我更新和分化成多种神经细胞类型的功能,而且被证明可以在移植入动物模型脑后成功分化为多巴胺神经元.目的:观察移植的骨髓间充质干细胞对锰中毒大鼠行为学以及脑内多巴胺神经元的影响.方法:通过向 SD 大鼠腹腔内注射四水氯化锰构建锰中毒大鼠模型,将锰中毒大鼠随机分为骨髓间充质干细胞组和对照组,分别向锰中毒大鼠右侧纹状体内移植5 μL第3代人骨髓间充质干细胞悬液或等量PBS.移植1个月后对大鼠进行行为学评分.通过免疫荧光观察骨髓间充质干细胞的分化情况,用ELISA检测大鼠右侧纹状体内多巴胺、脑源性神经营养因子、胶质细胞源性神经营养因子的含量.结果与结论:移植后观察到骨髓间充质干细胞组较对照组行为学评分显著下降.移植后的骨髓间充质干细胞组脑内可见人类特异性细胞核/酪氨酸羟化酶和人类特异性细胞核/胶质细胞源性酸性蛋白双标阳性细胞,而对照组脑内未见此两类双标阳性细胞;骨髓间充质干细胞组多巴胺、脑源性神经营养因子、胶质细胞源性神经营养因子含量与对照组接近.这表明骨髓间充质干细胞可以改善锰中毒大鼠的行为学,可以分化为多巴胺神经元和星形胶质细胞.%BACKGROUND: Studies have shown that bone marrow mesenchymal stem cells (BMSCs) can treat central nervous system diseases. BMSCs have the function of self-renewal and differentiation into a variety of neural cell types. BMSCs with self-renewal and multi-directional differentiation abilities can successfully differentiate into dopaminergic neurons after transplantation into an animal model. OBJECTIVE: To observe the effect of transplanted BMSCs on behavior and dopaminergic neurons in rats with manganese poisoning. METHODS: Rat models of manganese poisoning were constructed by intraperitoneal injection of MnCl2?4H2O into Sprague-Dawley rats. The model rats were then randomly divided into two groups, BMSCs and phosphate buffered solution (PBS) control group, and 5 μL of passage 3 human BMSCs suspension or equivalent PBS was transplanted into the right striatum of the manganese poisoning rats. One month after transplantation, the rats were subjected to behavioral assessment. The differentiation of BMSCs was observed by immunofluorescence. The contents of dopamine, brain-derived neurotrophic factor, glial cell-derived neurotrophic factor in the right striatum of rats were detected by ELISA. RESULTS AND CONCLUSION: The behavioral score of the BMSCs treated group was significantly lower than that of the PBS control group after transplantation (P < 0.05). Double-labeled positive cells for human-specific nuclear antigen/tyrosine hydroxylase (hNUC/TH) and human-specific nuclear antigen/glial cell-derived acidic protein (hNUC/GFAP) were observed in the BMSCs treated group after transplantation. Meanwhile, hNUC/TH and hNUC/GFAP double-labeled positive cells were undetected in the PBS control group after transplantation. The expression levels of dopamine, brain-derived neurotrophic factor, glial cell-derived neurotrophic factor in the BMSCs treated group were higher than those in the PBS control group. This suggests that BMSCs can improve the behavior of manganese poisoning rats and can differentiate into dopaminergic neurons and astrocytes.