黄芪甲苷对实验性自身免疫性脑脊髓炎小鼠T细胞免疫调节的影响  

Immunomodulatory effect of astragaloside IV on T cells of experimental autoimmune encephalomyelitis mice

作　　者：穆秉桃 于婧文[1] 刘春云[1] 郭敏芳[1] 孟涛 杨鹏伟 魏文悦 宋丽娟 尉杰忠[1] 马存根[1,3] Mu Bingtao;Yu Jingwen;Liu Chunyun;Guo Minfang;Meng Tao;Yang Pengwei;Wei Wenyue;Song Lijuan;Yu Jiezhong;Ma Cungen(Institute of Brain Science,Shanxi Datong University,Datong 037009,Shanxi Province,China;The Second People’s Hospital of Jincheng,Jincheng 048000,Shanxi Province,China;Research Center of Neurobiology,Shanxi University of Chinese Medicine/The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis,National Administration of Traditional Chinese Medicine,Jinzhong 030619,Shanxi Province,China;Department of Physiology,Shanxi Medical University,Taiyuan 030032,Shanxi Province,China)

机构地区：[1]山西大同大学脑科学研究所,山西省大同市037009 [2]晋城市第二人民医院,山西省晋城市048000 [3]山西中医药大学神经生物学研究中心/国家中医药管理局多发性硬化益气活血重点研究室,山西省晋中市030619 [4]山西医科大学生理学系,山西省太原市030032

出　　处：《中国组织工程研究》2024年第7期1057-1062,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学青年基金项目(82004028),项目负责人:宋丽娟;中国科学院遗传与发育生物学研究所分子发育生物学国家重点实验室开放课题(2020-MDB-KF-09),项目负责人:宋丽娟;山西省卫健委医学科技领军团队(2020TD05),项目负责人:马存根;山西中医药大学青年科学家培育项目(2021-PY-QN-09),项目负责人:宋丽娟;山西省黄芪资源产业化及产业国际化协同创新中心项目(HQXTCXZX2016-022),项目负责人:马存根;大同市应用基础研究计划项目(2020145),项目负责人:于婧文。

摘　　要：背景:多发性硬化初始阶段,中枢免疫细胞激活并释放大量炎症因子,引起白质脱髓鞘甚至累及灰质神经元。CD4^(+)T细胞不同亚群之间的分化平衡在实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)的病程进展中发挥着重要作用。课题组前期研究结果表明黄芪内有效成分黄芪甲苷能够调节EAE小鼠体内免疫反应,其是否对T细胞亚群分化具有调节作用尚未明确。目的:探究黄芪甲苷对EAE小鼠治疗效果及其对T细胞的免疫调控机制。方法:将C57BL/6雌性小鼠分为正常对照组、EAE疾病模型组和黄芪甲苷治疗组,每组8只,后2组使用髓鞘少突胶质细胞糖蛋白35-55(MOG35-55)制备EAE模型,免疫后第10-28天,黄芪甲苷治疗组以40 mg/(kg·d)灌胃给药。免疫当天至第28天,记录各组小鼠的体质量及临床评分;免疫后第28天取小鼠脊髓制成冰冻切片行苏木精-伊红染色、固蓝染色观察脊髓病理改变,流式细胞术检测脾脏T细胞亚群百分比,Western blot法检测脊髓组织中γ干扰素、白细胞介素17、白细胞介素6的蛋白表达,ELISA检测脾细胞上清液中γ干扰素、白细胞介素17、白细胞介素6、白细胞介素4水平。结果与结论:(1)与EAE疾病模型组相比,黄芪甲苷治疗能够减少EAE小鼠体质量丢失(P<0.05),缓解临床症状(P<0.05),减轻脊髓炎症细胞浸润及髓鞘脱失病理改变(分别为P<0.01和P<0.05);(2)与EAE疾病模型组相比,黄芪甲苷治疗可抑制表达γ干扰素和白细胞介素17的CD4^(+)T细胞亚群比例(分别为P<0.001和P<0.001),上调表达白细胞介素10和转化生长因子β的CD4^(+)T细胞亚群百分比(分别为P<0.001和P<0.01);(3)黄芪甲苷可下调脊髓和脾脏中γ干扰素(分别为P<0.05和P<0.01)、白细胞介素17(分别为P<0.05和P<0.05)、白细胞介素6(分别为P<0.05和P<0.05)的表达,上调脾脏中抑炎因子白细胞介素4的表达(P<0.01);(4)结果说明,黄芪甲苷可以减轻EAE�BACKGROUND:In the initial stage of multiple sclerosis,central immune cells activate and release a large number of inflammatory factors,causing white matter demyelination and even involving gray matter neurons.The equilibrium of differentiation between different subsets of CD4^(+)T cells plays an important role in the progression of experimental autoimmune encephalomyelitis.The previous results of the research group showed that the active ingredient astragalus glycoprotein in astragalus can regulate the immune response in experimental autoimmune encephalomyelitis mice,and whether it has a regulatory effect on the differentiation of T cell subsets has not been determined.OBJECTIVE:To explore the therapeutic effects and immune regulatory mechanisms of astragaloside IV on experimental autoimmune encephalomyelitis mice.METHODS:Female C57BL/6 mice were divided into the normal control group,experimental autoimmune encephalomyelitis disease model group,and astragaloside IV treatment group(n=8 per group).Myelin oligodendrocyte glycoprotein peptides 35-55 were used for experimental autoimmune encephalomyelitis model induction in the last two groups.On day 10 to 28 after immunization,the astragaloside IV treatment group was treated with 40 mg/kg per day astragaloside IV intragastrically.Body weight and clinical scores of mice in each group were recorded from the immunization day to the 28th day.On the 28th day after immunization,the mouse spinal cord was taken and made into frozen sections for hematoxylin-eosin staining and Lux fast blue staining to observe pathological changes in the spinal cord.Percentage of splenic T cell subsets was detected using flow cytometry.Western blot assay was used to determine the protein expression of interferon-γ,interleukin-17 and interleukin-6 in the spinal cord.Levels of interferon-γ,interleukin-17,interleukin-6 and interleukin-4 in supernatants of cultured splenocytes were determined by ELISA.RESULTS AND CONCLUSION:(1)Compared with the experimental autoimmune encephalomyelitis disease m

关 键 词：中枢神经系统 多发性硬化 实验性自身免疫性脑脊髓炎 髓鞘少突胶质细胞糖蛋白35-55 黄芪甲苷 T细胞 炎症 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R744.51