Stem Cells and Biotherapy Engineering Research Center of Henan, National Joint Engineering Laboratory of Stem Cells and Biotherapy, School of Life Science and Technology, Xinxiang Medical University, Xinxiang, 453003, China.
Department of Neurology, The First Affiliated Hospital of Xinxiang Medical University, Xinxiang, 45003, China.
Stem Cell Res Ther. 2023 Apr 13;14(1):85. doi: 10.1186/s13287-023-03330-7.
Neuroinflammation is closely related to the development of Parkinson's disease (PD). Because of the extensive sources, non-invasive and periodical collection method, human menstrual blood-derived endometrial stem cells (MenSCs) have been explored as a promising tool for treatment of PD. This study aimed to investigate if MenSCs could inhibit neuroinflammation in PD rats by regulating M1/M2 polarization and to excavate the underlying mechanisms.
MenSCs were co-cultured with 6-OHDA-exposed microglia cell lines. Then the morphology of microglia cells and the level of inflammatory factors were assessed by immunofluorescence and qRT-PCR. After MenSCs were transplanted into the brain of PD rats, animal motor function, the expression of tyrosine hydroxylase, and the level of inflammatory factors in the cerebrospinal fluid (CSF) and serum were detected to evaluate the therapeutic potential of MenSCs. Meanwhile, the expression of M1/M2 phenotype related genes was detected by qRT-PCR. One protein array kit containing 1000 kinds of factors was used to detect the protein components in the conditioned medium of MenSCs. Finally, bioinformatic analysis was performed to analyze the function of factors secreted by MenSCs and the signal pathways involved in.
MenSCs could suppress 6-OHDA-induced microglia cell activation and significantly decrease inflammation in vitro. After transplantation into the brain of PD rats, MenSCs improved animal motor function, which was indicated by the increased movement distance, ambulatory episodes, exercise time on the rotarod, and less contralateral rotation. Additionally, MenSCs reduced the loss of dopaminergic neurons and down-regulated the level of pro-inflammatory factors in the CSF and serum. Moreover, q-PCR and WB results showed the transplantation of MenSCs significantly down-regulated the expression of M1 phenotype cell markers and meanwhile up-regulated the expression of M2 phenotype cell markers in the brain of PD rats. 176 biological processes including inflammatory response, negative regulation of apoptotic process, and microglial cell activation were enriched by GO-BP analysis. 58 signal pathways including PI3K/Akt and MAPK were enriched by KEGG analysis.
In conclusion, our results provide preliminary evidence for the anti-inflammation capacity of MenSCs by regulating M1/M2 polarization. We firstly demonstrated the biological process of factors secreted by MenSCs and the signal pathways involved in using protein array and bioinformatic analysis.
神经炎症与帕金森病（PD）的发展密切相关。由于来源广泛、非侵入性和周期性采集方法，人类月经血源性子宫内膜干细胞（MenSCs）已被探索作为治疗 PD 的一种有前途的工具。本研究旨在探讨 MenSCs 是否可以通过调节 M1/M2 极化来抑制 PD 大鼠的神经炎症，并挖掘其潜在机制。
将 MenSCs 与 6-OHDA 暴露的小胶质细胞系共培养。然后通过免疫荧光和 qRT-PCR 评估小胶质细胞的形态和炎症因子水平。将 MenSCs 移植到 PD 大鼠脑内后，检测动物运动功能、酪氨酸羟化酶表达以及脑脊液（CSF）和血清中炎症因子水平，以评估 MenSCs 的治疗潜力。同时，通过 qRT-PCR 检测 M1/M2 表型相关基因的表达。使用包含 1000 种因子的蛋白质芯片试剂盒检测 MenSCs 条件培养基中的蛋白成分。最后，通过生物信息学分析分析 MenSCs 分泌的因子的功能及涉及的信号通路。
MenSCs 可抑制 6-OHDA 诱导的小胶质细胞活化，并显著降低体外炎症反应。移植到 PD 大鼠脑内后，MenSCs 改善了动物运动功能，表现为运动距离增加、运动次数增加、旋转棒上的运动时间增加、对侧旋转减少。此外，MenSCs 减少了多巴胺能神经元的丢失，并下调了 CSF 和血清中促炎因子的水平。此外，q-PCR 和 WB 结果表明，MenSCs 移植显著下调了 PD 大鼠脑中 M1 表型细胞标志物的表达，同时上调了 M2 表型细胞标志物的表达。GO-BP 分析富集了 176 个生物学过程，包括炎症反应、凋亡过程的负调控和小胶质细胞激活。KEGG 分析富集了 58 个信号通路，包括 PI3K/Akt 和 MAPK。
总之，我们的结果提供了 MenSCs 通过调节 M1/M2 极化发挥抗炎作用的初步证据。我们首次使用蛋白质芯片和生物信息学分析，证明了 MenSCs 分泌的因子的生物学过程和涉及的信号通路。
