Department of Geriatrics, The Second Affiliated Hospital, Nanjing Medical University , Nanjing, China.
Department of Neurology, The Affiliated Jiangning Hospital of Nanjing Medical University , Nanjing, China.
Neurol Res. 2021 Mar;43(3):199-209. doi: 10.1080/01616412.2020.1834775. Epub 2020 Oct 19.
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) amyotrophic lateral sclerosis (ALS) mice.
The BDNF gene was transfected into hUC-MSC-derived motor neurons by the lentivirus-mediated method. hSOD1 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.
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.
Transplantation of BDNF-overexpressing hUC-MSC-derived motor neurons can improve motor performance and prolong the survival of hSOD1 mice. Combining stem cell-derived motor neurons with BDNF might provide a new therapeutic strategy for ALS.
研究脑源性神经营养因子（BDNF）过表达的人脐带来源间充质干细胞（hUC-MSC）衍生运动神经元对人铜锌超氧化物歧化酶 1（hSOD1）肌萎缩侧索硬化症（ALS）小鼠的有益作用。
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。将 hSOD1 小鼠分为 ALS、ALS/MN 和 ALS/MN-BDNF 组，分别鞘内给予磷酸盐缓冲液（PBS）、运动神经元或过表达 BDNF 的运动神经元。对照组包括接受 PBS 治疗的非转基因野生型同窝仔鼠。移植后 1 个月，通过转棒试验评估小鼠的运动功能，然后分离腰椎增大区，通过 Western blot 检测 hSOD1 和 BDNF 的表达，通过免疫荧光法检测胆碱乙酰转移酶（ChAT）、同源盒蛋白 9（HB9）、主要组织相容性复合体 I（MHCI）和微管相关蛋白-2（MAP-2）的表达。
移植后，ALS/MN-BDNF 和 ALS/MN 组小鼠的跌倒潜伏期和存活时间均长于 ALS 组（P < 0.01 比 P < 0.05），前者的改善程度大于后者。然而，细胞移植并未延迟疾病发作。在 ALS/MN-BDNF 和 ALS/MN 组的腰椎增大区，hSOD1 的表达略有减少，但无统计学意义（P > 0.05），但 BDNF、ChAT 和 HB9 的表达以及 MHCI 和 MAP-2 的共表达明显大于 ALS 组（P < 0.01），前者的差异也更为显著。
移植过表达 BDNF 的 hUC-MSC 衍生运动神经元可改善运动性能并延长 hSOD1 小鼠的存活时间。将干细胞衍生的运动神经元与 BDNF 结合可能为 ALS 提供新的治疗策略。
