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人脐带间充质干细胞来源外泌体可调控巨噬细胞的极化

Regulatory effect of human umbilical cord mesenchymal stem cells-derived exosomes on macrophage polarization

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【作者】 宋玉仙张东亚许玉君侯亚义泥艳红

【Author】 Song Yuxian;Zhang Dongya;Xu Yujun;Hou Yayi;Ni Yanhong;Nanjing Stomatological Hospital, Medical School of Nanjing University;Immunology Laboratory and Jiangsu Key Laboratory of Molecular Medicine, Medical School of Nanjing University;

【通讯作者】 宋玉仙;

【机构】 南京大学医学院附属口腔医院南京市口腔医院南京大学医学院免疫学实验室和江苏省医学分子技术重点实验室

【摘要】 背景:间充质干细胞来源外泌体具有与母细胞类似的修复组织损伤的作用,但机制尚未完全阐明。巨噬细胞在组织损伤修复过程中扮演重要角色。因此,探讨间充质干细胞来源外泌体对巨噬细胞极化的调控作用,将有助于阐明其组织修复的机制。目的:提取人脐带间充质干细胞来源外泌体,并探讨其对巨噬细胞极化的影响。方法:采用超速离心法提取人脐带间充质干细胞分泌的外泌体,分别采用透射电镜观察其形态、纳米颗粒追踪法检测粒径、蛋白印迹法检测标志蛋白表达;然后用得到的外泌体处理小鼠骨髓巨噬细胞,探讨其对巨噬细胞极化的调控作用。结果与结论:①成功提取人脐带间充质干细胞来源外泌体,其形态呈经典的囊泡样结构,直径(92.0±34.1) nm,表达外泌体标志蛋白CD63和Alix;②该外泌体能显著降低脂多糖诱导的巨噬细胞M1型活化标志肿瘤坏死因子α和一氧化氮合酶水平,而提高M2型活化标志白细胞介素10和精氨酸酶1水平;③结果可见,人脐带间充质干细胞来源外泌体具有调控巨噬细胞向M2型极化的作用,进而发挥组织修复的功能。

【Abstract】 BACKGROUND: Accumulating evidence has suggested that mesenchymal stem cells-derived exosomes have the similar tissue repair activity compared to the mother cells. However, the underlying mechanisms are not yet illuminated. Since macrophages play pivotal roles during tissue repair process, we investigated the regulatory effect of mesenchymal stem cells-derived exosomes on macrophages, aiming to better elucidate its mechanism involved in tissue repair.OBJECTIVE: To isolate exosomes from human umbilical cord-derived mesenchymal stem cells, and to evaluate their regulatory effect on macrophage polarization.METHODS: Exosomes were isolated from the culture supernatants of human umbilical cord-derived mesenchymal stem cells by the differential centrifugation method, and then characterized by their morphology, size distribution and protein markers using transmission electron microscopy, nanoparticle tracking analysis and western blot assay, respectively. Furthermore, their regulatory effects on M1/M2 macrophage polarization were evaluated in mouse bone marrow-derived macrophages.RESULTS AND CONCLUSION: Exosomes were successfully isolated from human umbilical cord-derived mesenchymal stem cell supernatants with a cup-shaped canonical morphology,(92.0±34.1) nm in diameter, and protein markers of CD63 and Alix. We determined that these exosomes could significantly reduce lipopolysaccharide-induced activation of M1 markers tumor necrosis factor-α and inducible nitric oxide, while upregulating M2 markers interleukin-10 andarginase-1. These findings suggest that exosomes from human umbilical cord-derived mesenchymal stem cells can regulate macrophage polarization to M2 phenotype, benefiting tissue repairing.

【基金】 国家自然科学基金(81702680),项目负责人:宋玉仙;南京市医学科技发展项目(YKK17138),项目负责人:宋玉仙~~
  • 【文献出处】 中国组织工程研究 ,Chinese Journal of Tissue Engineering Research , 编辑部邮箱 ,2019年13期
  • 【分类号】R329.2
  • 【被引频次】7
  • 【下载频次】967
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