黄芪总皂苷对实验性自身免疫性脑脊髓炎小鼠的神经保护及修复机制研究  

Neuroprotective effect and repair mechanism of total saponins of astragalus on experimental autoimmune encephalomyelitis in mice

作　　者：刘建春 张红珍 李俊莲 樊慧杰 柴智 尉杰忠[2] 于婧文[2] 康东坤 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;KANG Dong-kun;XIAO Bao-guo;MA Cun-gen(Research Center of Neurobiology,Shanxi University of Chinese Medicine,Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of National Administration of Traditional Chinese Medicine,Jinzhong 030619,China;Institute of Brain Science,Shanxi Datong University,Shanxi Key Laboratory of Inflammatory Neurodegenerative Diseases,Datong 037009,China;Institute of Neurology,Huashan Hospital,Fudan University,Shanghai 200025,China)

机构地区：[1]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,晋中030619 [2]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 [3]复旦大学附属华山医院神经病学研究所,医学神经生物学国家重点实验室,上海200025

出　　处：《中华中医药杂志》2021年第11期6700-6704,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.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(No.HQXTCXZX2016-020);山西中医药大学科技创新能力培育计划项目(No.2019PY-027,No.2020PY-YC-25);山西省中医药管理局项目(No.2020ZYYC007);国家中医药管理局多发性硬化益气活血重点研究室,山西省卫健委医学科技领军团队(No.2020TD05)。

摘　　要：目的:探讨黄芪总皂苷(AST)对实验性自身免疫性脑脊髓炎(EAE)小鼠的保护作用,并探讨其神经修复的机制。方法:将18只C57BL/6雌性小鼠采用髓鞘少突胶质细胞糖蛋白35-55肽段(MOG35-55)诱导建立EAE模型,随机分为3组:EAE对照组,AST高、低剂量组。造模的第3天起,进行注射用药,持续25d。EAE对照组腹腔注射PBS;AST高、低剂量组分别腹腔注射黄芪总皂苷溶液80、40mg·kg^(-1)·d^(-1);各组小鼠每次给药0.2mL/只,持续记录体质量变化、临床评分和小鼠症状体征。HE染色检测脊髓炎性细胞浸润,髓鞘染色观察髓鞘脱失情况。ELISA法检测外周血和脾中白细胞介素(IL)-1β和IL-10的表达量。Western Blot法检测中枢神经组织中神经营养因子BDNF、GDNF和GAP-43的表达。结果:AST高、低剂量组均能明显推迟EAE起病时间,缓解EAE发病程度,AST高剂量的治疗效果优于低剂量;AST可抑制脊髓炎性细胞浸润,减少髓鞘脱失;血清和脾上清中IL-1β表达降低(P<0.01),IL-10表达升高(P<0.05);AST治疗可上调BDNF和GAP-43的表达。结论:AST干预可减轻EAE的发病程度,作用机制可能为调节细胞因子的表达和改善局部神经组织生长的微环境,促进神经营养类物质的表达。Objective: To explore the protective effect of total saponins of astragalus(AST) on experimental autoimmune encephalomyelitis(EAE) mice and to explore the mechanism of nerve repair. Methods: Eighteen C57BL/6 female mice were induced by myelin oligodendrocyte glycoprotein 35-55(MOG35-55) peptide to establish EAE model, and were randomly divided into three groups: EAE control group, high and low dose of AST group. After 3 days of modeling, the mice were injected with drug till 25 days. The EAE control group was injected with phosphate buffer saline(PBS) through abdominal cavity;the AST high-dose group and the AST low-dose group were injected with AST 80, 40 mg·kg^(-1)·d^(-1) into the abdominal cavity respectively.Every mice in each group were given 0.2 mL/d Body mass, clinical scores, and signs and symptoms of the mice were recorded every other day. Inflammatory cell infiltrations of spinal cord were observed by HE staining Myelin staining observes the demyelination situation. The expression of IL-1β and IL-10 in peripheral blood and spleen supernatant were detected by ELISA.Western blot was used to detect the expression of neurotrophic factors BDNF, GDNF and GAP-43 in central nervous system.Results: The high-dose and low-dose of AST groups can significantly delay the onset of EAE and alleviate the incidence of EAE,and the treatment effect of high-dose of AST is better than that of low-dose AST. AST inhibited the inflammatory cell infiltration and demyelination in the nervous centralis. The expression of IL-1β in serum and spleen supernatant was decreased(P<0.01), and the expression of IL-10 was increased(P<0.05). AST treatment up-regulated the expression of BDNF and GAP-43. Conclusion:AST intervention can relieve the clinical symptoms of EAE mice, AST may regulate the expression of cytokines, the improvement of the local nerve growth microenvironment, and the promotion of the expression of NTFs.

关 键 词：实验性自身免疫性脑脊髓炎 黄芪总皂苷 细胞因子 神经营养因子 神经保护 神经修复 

分 类 号：R285.5[医药卫生—中药学]