黄芪多糖对实验性自身免疫性脑脊髓炎小鼠脾细胞的免疫调节作用研究  

Immunoregulatory effect of astragalus polysaccharide on spleen cells of experimental autoimmune encephalomyelitis mice

作　　者：刘建春 张红珍 李俊莲 樊慧杰 柴智 尉杰忠[3] 于婧文[3] 肖保国 马存根 LIU Jian-Chun;ZHANG Hong-Zhen;LI Jun-Lian;FAN Hui-Jie;CHAI Zhi;YU Jie-Zhong;YU Jing-Wen;XIAO Bao-Guo;MA Cun-Gen(Research Center of Neurobiology,Shanxi University of Traditional Chinese Medicine,Jinzhong 030619,China)

机构地区：[1]山西中医药大学神经生物学研究中心,晋中030619 [2]国家中医药管理局多发性硬化益气活血重点研究室,晋中030619 [3]山西大同大学脑科学研究所,中枢神经炎症变性疾病新药创制省市共建山西省重点实验室培育基地,大同037009 [4]复旦大学附属华山医院神经病学研究所,上海200025

出　　处：《中国免疫学杂志》2021年第14期1701-1705,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金2014年面上项目(81473577);山西中医药大学中西医结合二级学科,黄芪资源产业化及产业国际化协同创新中心项目(HQXTCXZX2016-020);山西中医药大学科技创新能力培育计划项目(2019PY-027);山西中医药大学科技创新能力培育计划山西药茶的研制与开发项目(2020PY-YC-25);山西省中医药管理局项目(No.2020ZYYC007)。

摘　　要：目的:探讨黄芪多糖(APS)对实验性自身免疫性脑脊髓炎(EAE)小鼠脾细胞的免疫调节作用。方法:C57BL/6雌性小鼠腹腔注射髓鞘少突胶质细胞糖蛋白35-55(MOG_(35-55))建立EAE模型,免疫后的第9天取脾,制备脾细胞,和/无APS培养48 h后,流式细胞术检测脾中CD4^(+)LT淋巴细胞表型及M1型、M2型巨噬细胞表型的表达情况;ELISA法测定脾上清中干扰素γ(IFN-γ)、IL-12及IL-10的水平;CCK-8法检测脾细胞增殖情况;用CFSE荧光标记,和/无APS培养脾细胞72 h后,检测脾细胞增殖情况。结果:与EAE组相比,APS高、低剂量均可使脾细胞增殖率明显上升(P<0.01);APS高剂量能显著增加CD25^(+)L、TGF-β^(+)L的CD4^(+)LT细胞亚群的比例,下调IL-12^(+)L的CD4^(+)LT细胞的表达(P<0.01)。APS低剂量可显著增加M2型巨噬细胞表面标志CD206的表达(P<0.001),使脾细胞上清液中IL-10分泌增高,IL-12分泌降低(P<0.05);APS高、低剂量均可显著抑制M1型巨噬细胞表面标志物CD16/32^(+)L的表达(P<0.001)。结论:APS可通过调节抗炎和致炎免疫细胞的比例,调节细胞因子的含量,发挥免疫调节作用,提示APS具有治疗EAE的潜能。Objective:To investigate the immunomodulatory effects of astragalus polysaccharides(APS)on spleen cells in experimental autoimmune encephalomyelitis(EAE)mice.Methods:C57 BL/6 female mice were induced to establish an EAE model with myelin oligodendrocyte glycoprotein 35-55(MOG35-55).Spleen was taken on day 9 after immunization and splenocytes were prepared,with/without APS was cultured for 48 h,flow cytometry was performed to detect the expression of CD4^(+)LT lymphocyte phenotype and M1/M2 macrophages phenotype in the spleen.The levels of IFN-γ,IL-12 and IL-10 in spleen supernatant were determined by ELISA.The proliferation of splenocytes was detected by CCK-8 method.Splenocytes with CFSE fluorescent label,with/without APS was cultured for 72 h,flow cytometry was performed to detect proliferation of splenic cells.Results:Compared with the EAE group,the proliferation rate of splenic cells increased significantly with both high and low doses of APS(P<0.01).High dose of APS significantly increased the proportion of CD4^(+)LT cell subsets of CD25+Land TGF-β^(+) L,and down-regulated the expression of CD4^(+)LT cells of IL-12^(+)L(P<0.01).The low dose of APS significantly increased the expression of M2 macrophage surface marker CD206(P<0.001),resulting in increased IL-10 secretion and decreased IL-12 secretion in splenic supernatant(P<0.05).Both high and low doses of APS significantly inhibited the expression of M1 macrophages surface marker CD16/32^(+) L(P<0.001).Conclusion:APS by adjusting the proportion of proinflammatory and anti-inflammatory immune cells,and adjusting the content of cytokines,suggesting that astragalus polysaccharides has the potential to treat EAE.

关 键 词：黄芪多糖 实验性自身免疫性脑脊髓炎 脾细胞 CD4+T细胞 巨噬细胞 

分 类 号：R285[医药卫生—中药学]