补阳还五汤对EAE小鼠免疫调节的作用  

Immunoregulatory effect of Buyang Huanwu Decoction on experimental autoimmune encephalomyelitis mice

作　　者：王晓庆[1] 田倩倩[1] 李艳花[2] 刘建春[1] 刘春云[2] 杨春彦[1] 肖保国[3] 尉杰忠[2] 马存根[1] WANG Xiao-Qing TIAN Qian-Qian LI Yan-Hua LIU Jian-Chun LIU Chun-Yun YANG Chun-Yan XIAO Bao-Guo YU Jie-Zhong MA Cun-Gen(＂ 2011 ＂ Collaborative Innovation Center/Research Center of Neurobiology, Shanxi University of Traditional Chinese Medicine, Taiyuan 030024, China)

机构地区：[1]山西中医学院"2011"协同创新中心/神经生物学研究中心,太原030024 [2]山西大同大学脑科学研究所,大同037009 [3]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》2017年第1期52-57,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金2014年面上项目(81473577);山西省自然科学基金(2013011052-4);山西省国际科技合作项目(2013081058);山西省回国留学人员重点科研资助项目(2014-重点7);山西中医学院“2011”培育计划项目(2011PY-1)

摘　　要：目的:探讨补阳还五汤对实验性自身免疫性脑脊髓炎的抗炎治疗效果和免疫调节机制。方法:雌性C57BL/6小鼠采用MOG35-55多肽诱导建立EAE模型后,随机分为生理盐水组、补阳还五汤组,每组13只。造模的第3天给药,生理盐水组给予生理盐水25 ml/kg灌胃,补阳还五汤组给予补阳还五汤生药量50 g/kg灌胃,共治疗25 d。每天记录临床症状评分和体重变化。HE染色检测脊髓组织炎细胞浸润,髓鞘染色观察髓鞘脱失情况,免疫荧光技术检测脾脏ROCKⅡ表达。流式细胞术检测脾脏CD4^+T细胞亚群变化。Western blot法检测脊髓TLR4、Myd88、NF-κB、COX-2、ROCKⅡ和脑ROCKⅡ的表达。结果:与生理盐水组比较,补阳还五汤治疗后明显降低EAE小鼠神经功能评分(P<0.001),抑制中枢神经系统炎细胞浸润(P<0.05),减轻髓鞘脱失(P<0.05),增加CD25^+(P<0.05)、IL-10^+(P<0.05)、TGF-β^+(P<0.01)、IFN-γ^+(P<0.05)CD4^+T细胞的比例;抑制外周和中枢ROCKⅡ的表达(P<0.05);减少脊髓TLR4、Myd88、NF-κB、COX-2的表达(P<0.05)。结论:补阳还五汤可以通过抑制ROCKⅡ/TLR4/NF-κB炎性通路和调节外周T细胞亚群比例发挥其抗炎和免疫调节作用。Objective： To explore the anti-inflammatory therapeutic effect and possible immunoregulatory mechanism of Buyang Huanwu Decoction（ BYHWD） on the development of experimental autoimmune encephalomyelitis（ EAE）. Methods： Female C57 BL /6 mice were immunized subcutaneously with myelin oligodendrocyte glycoprotein peptides（ MOG35-55）,and randomly divided into saline group,BYHWD group,with 13 mice in each group. At the 3th day,25 ml / kg of saline was orally given to each mouse of saline group,50 g / kg ig of crude BYHWD was orally given to each mouse in BYHWD group for 25 days. Clinical score and body mass were recorded every day. Inflammatory cell infiltrations of spinal cord were observed by HE staining Myelin staining observes the demyelination situation. And the expression of ROCKⅡ in spleen was detected by immunofluorescence staining. The subtypes of CD4~＋T cells were analyzed by flow cytometry. Western blot was used to detect the expression of TLR4,Myd88,NF-κB,COX-2,ROCKⅡ in spinal cord and ROCKⅡ in brain. Results： The neurologicalscore significantly decreased in EAE mouse of BYHWD group compared with the saline group（ P〈0. 001）. BYHWD inhibited the inflammatory cell infiltration and demyelination in the nervous centralis（ P〈0. 05）. The treatment of BYHWD effectively reduced the increased the proportion of CD25~＋（ P〈0. 05）,IL-10~＋（ P〈0. 05）,TGF-β~＋（ P〈0. 01）,IFN-γ~＋（ P〈0. 05） CD4~＋T cells,and inhibited the expression of peripheral and central ROCKⅡ（ P〈0. 05）; BYHWD reduced the expression of TLR4,My D88,NF-κB,COX-2 in spinal cord（ P〈0. 05）. Conclusion： BYHWD can exert anti-inflammatory and immune regulation effect by the inhibition of ROCKⅡ/ TLR4 / NF-κB signaling pathway and regulation of the proportion of peripheral T cell subsets.

关 键 词：补阳还五汤 实验性自身免疫性脑脊髓炎 ROCKⅡ/TLR4/NF-κB炎性通路 

分 类 号：R285[医药卫生—中药学]