Research and Experimental Development Department, Asia Stem Cell Regenerative Pharmaceutical Co. Ltd, Shanghai, China.
Front Immunol. 2023 Jan 13;13:1095469. doi: 10.3389/fimmu.2022.1095469. eCollection 2022.
This study explored the underlying therapeutic mechanism of human umbilical cord mesenchymal stem cells (hUCMSCs) for ischemic stroke (IS), and determined the optimal administration time windows and dose-effect relationship.
The levels of SDF-1α, IL-10, IL-6, TNF-α, BDNF, IL-1β, and VEGF-A factors in serum and brain tissue lysate were measured by ELISA. The pathological status of brain tissues was evaluated by Hematoxylin-Eosin (HE) staining, and apoptosis of nerve cells was detected by tunel. The protein expression of CXCR-4, NeuN, and Nestin in the brain tissues was assessed through immunofluorescence. The balance beam, forelimb muscle strength, and limb placement were tested on MCAO rats at different time points and doses. The infarct area of the rat brain tissues was measured at the end of the experiment.
The hUCMSC treatment during the acute phase of MCAO significantly reduced the secretion of IL-6, TNF-α, IL-1β but increased IL-10 in serum, and the levels of SDF-α and BDNF in serum and brain tissues lysate were also increased. The pathological results showed that there were more neurons in the treatment group compared to the model group. Immunofluorescence assays showed that the expression of CXCR4、Nestin、NeuN was relatively higher than that in the model group. The d4 and d7 treatment significantly improves the motor function, promotes the recovery of forelimb muscle strength, increases the forelimb placement rate and reduces the scope of cerebral infarction, but the d14 treatment group has less therapeutic effect compared to the d4 and d7 treatment. The 2×10/kg treatment showed the best therapeutic effect, followed by the 1×10/kg treatment, and the worst is 0.5×10/kg treatment from the test of balance beam, forelimb muscle strength, limb placement and the infarct area of the rat brain tissues.
The hUCMSCs can inhibit the infiltration of inflammatory cells in the brain tissue, and promote the repair of brain tissue structure and function. Early intervention by injecting high-dose of hUCMSCs can significantly improve the recovery of neurological/motor function and reduce the size of cerebral infarction in rats.
本研究旨在探讨人脐带间充质干细胞（hUCMSCs）治疗缺血性脑卒中（IS）的潜在治疗机制，并确定最佳的给药时间窗和剂量效应关系。
采用 ELISA 法检测血清和脑组织裂解液中 SDF-1α、IL-10、IL-6、TNF-α、BDNF、IL-1β和 VEGF-A 因子的水平。通过苏木精-伊红（HE）染色评估脑组织的病理状态，通过 TUNEL 检测神经细胞凋亡。通过免疫荧光法评估脑组织中 CXCR-4、NeuN 和 Nestin 的蛋白表达。在不同时间点和剂量下对 MCAO 大鼠进行平衡梁、前肢肌力和肢体放置测试。实验结束时测量大鼠脑组织的梗死面积。
MCAO 急性期 hUCMSC 治疗显著降低了血清中 IL-6、TNF-α、IL-1β的分泌，但增加了血清和脑组织裂解液中 SDF-α和 BDNF 的水平。病理结果显示，治疗组的神经元数量多于模型组。免疫荧光检测显示，CXCR4、Nestin、NeuN 的表达相对高于模型组。d4 和 d7 治疗组明显改善运动功能，促进前肢肌力恢复，增加前肢放置率，减少脑梗死范围，但 d14 治疗组的治疗效果不如 d4 和 d7 治疗组。2×10/kg 治疗组表现出最佳的治疗效果，其次是 1×10/kg 治疗组，最差的是 0.5×10/kg 治疗组，从平衡梁、前肢肌力、肢体放置和大鼠脑组织梗死面积的测试结果可以看出。
hUCMSCs 可抑制脑组织中炎性细胞的浸润，并促进脑组织结构和功能的修复。早期注射高剂量 hUCMSCs 可显著改善神经/运动功能的恢复，减少大鼠脑梗死的体积。
