| 高 翔,楚佳奇,王家丰,赵名艳,黄 瑞,李 鹏.间充质干细胞来源外泌体对脑内小胶质细胞极化和炎症因子释放的影响及机制研究[J].,2021,(20):3809-3813 |
| 间充质干细胞来源外泌体对脑内小胶质细胞极化和炎症因子释放的影响及机制研究 |
| Effect and Mechanism of Mesenchymal Stem Cell-derived Exosomes on Microglia Polarization and Inflammatory Cytokine Release in Brain |
| 投稿时间：2021-03-10 修订日期：2021-04-13 |
| DOI：10.13241/j.cnki.pmb.2021.20.002 |
| 中文关键词: 间充质干细胞外泌体 小胶质细胞 炎症因子 |
| 英文关键词: Mesenchymal stem cells exosomes Microglia cells Inflammation cytokines |
| 基金项目:广东省医学科研基金项目（A2019463）；广东省自然科学基金面上项目（2018A030307036，2020A1515011565）；湛江市细胞治疗工程技术研究开发中心建设项目（2019A01032）；广东医科大学科研基金面上培育项目（GDMUM201808） |
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| 中文摘要: |
| 摘要 目的：探究间充质干细胞外泌体对脑内小胶质细胞极化和炎症因子释放的影响及其机制。方法：收集体外培养的间充质干细胞上清，超高速冷冻离心获取外泌体。采用纳米颗粒系统和透射电子显微镜分别检测外泌体粒径大小、形态结构和功能完整性。通过免疫荧光、ELISA和细胞流式等方式检测LPS刺激下，外泌体对BV2细胞的表型极化和炎症因子释放的影响。采用Western Blot法检测间充质干细胞外泌体对BV2细胞JAK1/STAT3通路活化的影响。结果：（1）间充质干细胞分泌的外泌体粒径大小主要介于40-100 nm，透射电镜显示外泌体形态呈典型膜性"杯盘"状结构；（2）流式结果表明，相比于对照组，LPS组能显著激活M1型小胶质细胞表面标志物CD11b和M2型小胶质细胞表面标志物CD206的表达，而经外泌体处理，CD11b的表达显著被抑制，CD206显著升高。同时ELISA结果证实，相比于LPS组，外泌体组分泌的促炎症因子（IL-1β、IL-6）和NO水平显著降低（P<0.05），抗炎因子（IL-10）显著升高 (P<0.05)；（3）间充质干细胞外泌体显著提高了BV2细胞JAK1/STAT3通路的磷酸化水平。结论：间充质干细胞外泌体通过激活JAK1/STAT3通路有效促进脑内小胶质细胞M1型向M2型极化。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effects of exosomes secreted by mesenchymal stem cells on microglia polarization and inflammatory cytokines release in brain and its possible mechanism. Methods: Exosomes were extracted and purified from the supernatant of mesenchymal stem cells cultured in vitro by ultrahigh speed cryogenic centrifugation. The particle size, morphological structure and functional integrity of exosomes were determined by nanoparticle tracking analysis and transmission electron microscopy. The effects of exosomes on the polarization stimulation function of BV2 cells and the release of inflammatory factors in the LPS-stimulated environment were detected by immunofluorescence, ELISA and flow cytometry. The activation of JAK1/STAT3 pathway in BV2 cells was detected by Western Blot. Results: (1) The granule size of exosomes secreted by mesenchymal stem cells was mainly 40-100 nm, and the transmission electron microscope showed a typical membrane "cup and plate" structure. (2) Flow cytometer results showed that compared with that in the control group, the expressions of M1-type microglial surface marker CD11b and M2-type microglial surface marker CD206 were significantly activated. In the stimulation of purified exosomes, the expression of CD11b was significantly inhibited, while CD206 was significantly activated. ELISA results showed that compared with that in the LPS group, the inflammatory cytokines (IL-1β, IL-6) and NO were significantly decreased (P<0.05), while anti-inflammatory cytokine IL-10 was significantly increased (P<0.05) in the exosomes group. (3) Mesenchymal stem cell-derived exosomes significantly increased the phosphorylation level of JAK1/STAT3 pathway in BV2 cells. Conclusion: Mesenchymal stem cell-derived exosomes can effectively promote M2-type polarization and release of anti-inflammatory factors of microglia in the brain by activating the JAK1/STAT3 pathway. |
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