ABSTRACT

Objective

To investigate the beneficial effect of brain-derived neurotrophic factor (BDNF) -overexpressing human umbilical cord mesenchymal stem cell (hUC-MSC)-derived motor neurons in the human Cu, Zn-superoxide dismutase1 (hSOD1)^G93A amyotrophic lateral sclerosis (ALS) mice.

Methods

The BDNF gene was transfected into hUC-MSC-derived motor neurons by the lentivirus-mediated method. hSOD1^G93A mice were assigned to the ALS, ALS/MN, and ALS/MN-BDNF groups, and intrathecally administrated phosphate-buffered saline (PBS), motor neurons, or motor neurons overexpressing BDNF, respectively. The control group included non-transgenic wild-type littermates administrated PBS. One month after transplantation, the motor function of the mice was assessed by the rotarod test, and the lumbar enlargements were then isolated to detect the expression of hSOD1 and BDNF by western blotting, and the expression of choline acetyltransferase (ChAT), homeobox protein 9 (HB9), major histocompatibility complex I (MHCI) and microtubule-associated protein-2 (MAP-2) by immunofluorescence assay.

Results

After transplantation, mice in the ALS/MN-BDNF and ALS/MN groups both exhibited longer latency to fall and longer survival than those in the ALS group (P < 0.01 vs. P < 0.05), and the improvement was more significant in the former than in the latter. However, cell transplantation did not delay disease onset. In the lumbar enlargements of the ALS/MN-BDNF and ALS/MN groups, the expression of hSOD1 was slightly reduced without statistical significance (P > 0.05), but the expression of BDNF, ChAT and HB9, and the co-expression of MHCI and MAP-2 were significantly greater than in the ALS group (P < 0.01), with the differences also being more prominent in the former group than in the latter.

Conclusions

Transplantation of BDNF-overexpressing hUC-MSC-derived motor neurons can improve motor performance and prolong the survival of hSOD1^G93A mice. Combining stem cell-derived motor neurons with BDNF might provide a new therapeutic strategy for ALS.

中文翻译：

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BDNF 过度表达的人脐带间充质干细胞衍生的运动神经元可改善肌萎缩侧索硬化小鼠的运动功能并延长其存活时间

摘要

客观的

研究脑源性神经营养因子 (BDNF) 过表达人脐带间充质干细胞 (hUC-MSC) 源性运动神经元对人铜、锌超氧化物歧化酶 1 (hSOD1) ^G93A肌萎缩侧索硬化 (ALS) 的有益影响老鼠。

方法

通过慢病毒介导的方法将BDNF基因转染到hUC-MSC衍生的运动神经元中。hSOD1 ^G93A小鼠被分配到 ALS、ALS/MN 和 ALS/MN-BDNF 组，并分别鞘内注射磷酸盐缓冲盐水 (PBS)、运动神经元或过表达 BDNF 的运动神经元。对照组包括给予PBS的非转基因野生型同窝仔。移植后1个月，用旋转棒试验评估小鼠的运动功能，然后分离腰部膨大，用蛋白质印迹法检测hSOD1和BDNF的表达，以及胆碱乙酰转移酶（ChAT）、同源框蛋白9的表达(HB9)、主要组织相容性复合物 I (MHCI) 和微管相关蛋白-2 (MAP-2) 通过免疫荧光测定。

结果

移植后，ALS/MN-BDNF和ALS/MN组小鼠均表现出较ALS组更长的跌倒潜伏期和更长的存活时间（P < 0.01 vs. P < 0.05），且改善更为显着。前者不如后者。然而，细胞移植并没有延迟疾病的发作。在ALS/MN-BDNF和ALS/MN组腰椎肿大中，hSOD1的表达略有降低，无统计学意义（P > 0.05），但BDNF、ChAT和HB9的表达，以及M​​HCI的共表达和 MAP-2 均显着高于 ALS 组（P < 0.01），且前者的差异也较后者更为显着。

结论

移植过表达 BDNF 的 hUC-MSC 衍生的运动神经元可以改善运动性能并延长 hSOD1 ^G93A小鼠的存活时间。将干细胞衍生的运动神经元与 BDNF 相结合可能为 ALS 提供一种新的治疗策略。