摘要
目的探索干细胞外泌体在干细胞发挥免疫调节中的关键作用,探讨人脐带间充质干细胞外泌体在系统性红斑狼疮中的潜在应用价值。方法选取2022年1月至2023年1月河南省人民医院系统性红斑狼疮患者(均为女性)的外周血标本15份。提取并鉴定人脐带间充质干细胞和人脐带间充质干细胞外泌体。从符合评分标准的系统性红斑狼疮患者的外周血中分离单个核细胞和T细胞,分别与脐带间充质干细胞和脐带间充质干细胞外泌体共培养48 h。通过ELISA检测共培养上清中炎症因子水平,RT-qPCR检测共培养后细胞相关基因mRNA的表达、流式细胞术检测调节性T细胞增殖能力,蛋白质印迹检测相关蛋白表达水平。结果成功提取脐带间充质干细胞和脐带间充质干细胞外泌体;脐带间充质干细胞和脐带间充质干细胞外泌体与外周血单个核细胞共培养后,均明显促进系统性红斑狼疮患者外周血单个核细胞中调节性T细胞的增殖分化,而且外周血单个核细胞中抗炎细胞因子转化生长因子β1(transforming growth factor-beta1,TGF-β1)和白细胞介素-10(interleukin-10,IL-10)的表达水平明显升高,促炎细胞因子γ干扰素(interferon-gamma,IFN-γ)水平显著下降;脐带间充质干细胞和脐带间充质干细胞外泌体对单一T细胞中调节性T细胞的诱导分化作用不明显。结论人脐带间充质干细胞外泌体具有与人脐带间充质干细胞相似的抗炎和促调节性T细胞增殖的功能,而且避免了人脐带间充质干细胞移植出现的细胞逃逸、不良分化、免疫原性等安全性问题。人脐带间充质干细胞外泌体通过外周血单个核细胞中非T细胞间接促进调节性T细胞增殖,降低炎症水平,重塑外周血免疫平衡,在系统性红斑狼疮治疗中具有潜在的应用价值。
Objective To investigate the pivotal role of exosomes in the immunomodulatory effects of stem cells and to evaluate the potential therapeutic application of exosomes derived from umbilical cord mesenchymal stem cells in systemic lupus erythematosus(SLE).Methods Totally 15 peripheral blood samples from patients with systemic lupus erythematosus(all female)in Henan Provincial People's Hospital were selected from January 2022 to January 2023.Human umbilical cord mesenchymal stem cell(Hu-MSCs)and Human umbilical cord mesenchymal stem cell exosomes(Hu-MSC-exo)were extracted and identified.Peripheral blood mononuclear cells(PBMCs)and T cells were extracted from patients with SLE meeting the scoring criteria and co-cultured with Hu-MSCs or Hu-MSC-exo for 48h.Cytokine levels in the culture supernatant were measured by ELISA,mRNA expression of related genes was analyzed by RT-qPCR,cell proliferation was assessed by flow cytometry,and protein expression levels were evaluated by Western blot.Results Hu-MSCs and Hu-MSC-exo were successfully extracted.After co-culture with PBMCs revealed that Hu-MSC-exo significantly increased the expression of transforming growth factor-betal and interleukin-10,while reducing interferon-gamma expression.Additionally,Hu-MSC-exo promoted the proliferation and differentiation of regulatory T cells.When co-cultured with T cells,there was no significant change in the number of regulatory T cells.Conclusion Hu-MSC-exo exhibit anti-inflammatory properties and the ability to promote regulatory T cell proliferation,functions comparable to those of Hu-MSCs.Importantly,Hu-MSC-exo circumvent safety concerns associated with Hu-MSCs transplantation,such as cell escape,aberrant differentiation,and immunogenicity.Hu-MSC-exo facilitate regulatory T cell expansion indirectly via non-T cells within PBMCs,thereby reducing inflammatory responses and restoring immune homeostasis in peripheral blood.This mechanism highlights their promising therapeutic potential for systemic lupus erythematosus.
作者
李亚龙
安乐美
张婷婷
程朝飞
张永辉
申思宁
岳寒
LI Ya-long;AN Le-mei;ZHANG Ting-ting;CHENG Chao-fei;ZHANG Yong-hui;SHEN Si-ning;YUE Han(Henan Key Laboratory of Stem Cell Clinical Application and Key Technology,Stem Cell Research Center,Peoples Hospital of Zhengzhou University,Henan Provincial Peoples Hospital,Zhengzhou,Henan 450003,China;Department of Rheumatology and Immunology,Peoples Hospital of Zhengzhou University,Henan Provincial Peoples Hospital,Zhengzhou,Henan 450003,China;Center for Clinical Single-Cell Biomedicine,Peoples Hospital of Zhengzhou University,Henan Provincial Peoples Hospital,Zhengzhou,Henan 450003,China;Department of Thoracic Surgery,Henan Cancer Hospital,Zhengzhou,Henan 450003,China)
出处
《医药论坛杂志》
2025年第15期1569-1575,共7页
Journal of Medical Forum
基金
河南省科技攻关计划项目(232102311040,232102311073,242102311165)
河南省自然科学基金青年基金(232300421297)。
关键词
人脐带间充质干细胞
外泌体
系统性红斑狼疮
调节性T细胞
免疫调节
Human umbilical cord mesenchymal stem cell
Exosome
Systemic lupus erythematosus
Regulatory T cells
Immunomodulatory
