摘要
目的探究不同浓度骨髓间充质干细胞外泌体对巨噬细胞向M1型极化的影响。方法提取骨髓间充质干细胞外泌体,电子显微镜观察外泌体形态,并通过Western blot检测骨髓间充质干细胞外泌体特异性蛋白TSG101、CD63、Alix表达水平;将不同浓度骨髓间充质干细胞外泌体与巨噬细胞共培养,RT-qPCR检测巨噬细胞极化相关基因IL-6、iNOS、Arg-1、CD206等的表达。结果电镜检测显示,外泌体为杯口状结构;Western blot检测显示骨髓间充质干细胞外泌体表达特异性蛋白TSG101、CD63、Alix;外泌体内吞实验表明,在60μg/ml外泌体时,巨噬细胞对外泌体的内吞效率最高。RT-qPCR分析显示,60μg/ml外泌体处理使M1型巨噬细胞相关基因IL-6和iNOS表达升高最明显。结论骨髓间充质干细胞的外泌体可以诱导巨噬细胞向M1型巨噬细胞极化。
Objective To investigate the effect of bone marrow mesenchymal stem cell exosomes on macrophage polarization.Methods The exosomes of bone marrow mesenchymal stem cells were extracted,the morphology of exosomes was observed by electron microscope,and the expression levels of exosome specific proteins of bone marrow mesenchymal stem cells,TSG101,CD63 and Alix,were detected by Western blot;the exosomes of bone marrow mesenchymal stem cells at different concentrations were co-cultured with macrophages.The expression of macrophage polarization related genes,IL-6,iNOS,Arg-1 and CD206,were detected by RT-qPCR.Results Electron microscopic observation found that the exosomes were cup-shaped structure,and the specific proteins,TSG101,CD63 and Alix,were detectable by Western blot;The results of exocrine endocytosis experiment showed that macrophages had the highest endocytosis efficiency of exosomes at 60μg/ml of the exosomes.RT-qPCR results showed that 60μg/ml of the exosome treatment increased the expression of M1 macrophage related genes,IL-6 and iNOS,most significantly.Conclusion The exosomes of bone marrow mesenchymal stem cells can induce polarization of macrophages into M1 macrophages.
作者
刘文涛
王新月
杨毅
文诸缦
李云鹏
白生宾
Liu Wentao;Wang Xinyue;Yang Yi;Wen Zhuman;Li Yunpeng;Bai Shengbin(School of basic medicine,Xinjiang Medical University,Urumqi,830000;School of public health,Xinjiang Medical University,Urumqi,830000;The First Affiliated Hospital of Xinjiang Medical University,Urumqi,830000)
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2022年第3期232-238,共7页
Chinese Journal of Histochemistry and Cytochemistry
基金
自治区科技支疆项目:骨力学生物学协同创新平台(2020E0285)
国家自然科学基金项目(82160356)
新疆组织细胞工程重点实验室项目(ZZXB001)。
