羊膜间充质干细胞经PGE2/EP2途径调节类风湿关节炎患者T细胞免疫  

Amniotic mesenchymal stem cells regulate T cell immunity in patients with rheumatoid arthritis through PGE2/EP2 pathway

作　　者：彭淋珅 刘娟 李林艳 陈辉 王宇 于泓 吴雨曈 田梅 余丽梅 Peng Linshen;Liu Juan;Li Linyan;Chen Hui;Wang Yu;Yu Hong;Wu Yutong;Tian Mei;Yu Limei(Guizhou Province Key Laboratory of Cell Engineering,Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563003,China;Department of Rheumatology,Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563003,China;Collaborative Innovation Center of Tissue Damage Repair and Regenerative Medicine of Ministry of Education,Zunyi Medical University,Zunyi Guizhou 563003,China;Engineering Research Center for Guizhou Clinical Application of Stem Cells and Drug Research and Development,Zunyi Medical University,Zunyi Guizhou 563003,China;Department of Nephrology,Kweichow Moutai Hospital,Renhuai Guizhou 564500,China)

机构地区：[1]遵义医科大学附属医院贵州省细胞工程重点实验室,贵州遵义563003 [2]遵义医科大学附属医院风湿病科,贵州遵义563003 [3]遵义医科大学教育部组织损伤修复与再生医学协同创新中心,贵州遵义563003 [4]遵义医科大学贵州省干细胞临床应用与药物开发工程研究中心,贵州遵义563003 [5]贵州茅台医院肾内科,贵州仁怀564500

出　　处：《遵义医科大学学报》2024年第6期558-568,共11页Journal of Zunyi Medical University

基　　金：国家自然科学基金资助项目(NO:82060270);贵州茅台医院科研与人才培养资金项目(NO:MTky2022-40)。

摘　　要：目的探讨人羊膜间充质干细胞(AMSCs)调节类风湿性关节炎(RA)患者T细胞免疫的作用与涉及PGE2/EP2途径的机制。方法含RA患者自体血浆的培养基非接触共培养AMSCs与RA患者外周血来源T细胞,实验分为T细胞组、AMSCs组、共培养组和EP2组,流式细胞术检测T细胞亚群和炎症因子,ELISA法检测PGE2和IL-6,Western blot检测PI3K、Akt及其磷酸化蛋白水平。结果AMSCs降低RA患者T细胞增殖数量和Th17细胞比例,增高Treg细胞比例和Treg/Th17比值,减少共培养上清液IL-17含量,这些效应可被40 nmol/L的EP2受体拮抗剂AH6809所阻断。与AMSC组相比,共培养组细胞培养上清液中IL-1β、IL-6和IL-8的含量明显降低,EP2受体拮抗剂组与共培养组比较无明显差异。AMSCs增加T细胞p-PI3K蛋白水平,降低p-AKT蛋白水平,EP2受体拮抗剂只逆转AMSCs对p-AKT蛋白的下调。结论AMSCs抑制RA患者T细胞增殖、上调Treg/Th17和下调IL-17分泌的作用机制涉及激活PGE2/EP2途径。Objective To investigate the effect of human amniotic mesenchymal stem cells(AMSCs)on the regulation of T cell immunity in rheumatoid arthritis(RA)patients and the mechanism involved in the PGE2/EP2 pathway.Methods T lymphocytes from peripheral blood of RA patients and AMSCs were co-cultured without contact in culture media supplemented with RA autologous plasma.T lymphocyte sub-populations and inflammatory factors of the culture supernatant were detected by flow cytometry.PGE2 and IL-6 were detected by ELISA.PI3K,Akt,and their phosphorylated protein levels were detected by Western blot.Results AMSCs significantly decreased the proliferation of T lymphocytes of RA patients and the proportion of Th17 cells,while they increased the proportion of Treg cells and the ratio of Treg/Th17.Inflammatory factor IL-17 content was decreased in the co-culture supernatant.These effects could be blocked by the EP2 receptor antagonist AH6809 at 40 nmol/L.Compared with the AMSCs group,the levels of IL-1β,IL-6,and IL-8 in the cultured medium were significantly reduced,but there was no significant difference between the EP2 receptor antagonist group and the culture group.AMSCs significantly heightened the level of p-PI3K,and decreased the level of p-AKT protein in T lymphocytes.EP2 receptor antagonists could only reverse the down-regulation of p-AKT protein by AMSCs.Conclusion AMSCs inhibits T cells proliferation,up-regulates Treg/Th17,and down-regulates IL-17 secretion in RA patients.The mechanism is related to activation of PGE2/EP2 pathway.

关 键 词：羊膜间充质干细胞 类风湿性关节炎 T细胞 炎症因子 前列腺素E2受体 

分 类 号：R593.22[医药卫生—内科学]