摘要
目的探讨二氢丹参酮I(DHTS1)对双环己酮草酰二腙(cuprizone)诱导脱髓鞘保护作用的机制。方法DHTS1溶解于5 g/L羧甲基纤维素钠(CMC-Na)中,饲喂小鼠含2 g/L cuprizone的饲料制备脱髓鞘模型,给予DHTS1处理,实验动物随机分为CMC-Na正常组、CMC-Na联合cuprizone处理组、DHTS1联合cuprizone处理组。通过固蓝(LFB)组织化学染色、免疫荧光组织化学染色检测髓鞘碱性蛋白(MBP)和髓鞘蛋白脂(PLP)的表达判断髓鞘完整性,原位末端转移酶标记技术(TUNEL)检测细胞凋亡;免疫荧光组织化学染色检测小胶质细胞/巨噬细胞特异性蛋白钙离子结合接头蛋白分子1(Iba-1)、CD86、CD163的表达确定小胶质细胞极化情况。脂多糖(LPS)极化SIM-A9小胶质细胞,DHTS1处理,实验分为CMC-Na对照组、DHTS1对照组、CMC-Na联合LPS处理组和DHTS1联合LPS处理组。流式细胞术检测CD16/32、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)阳性细胞比例。结果与CMC-Na联合cuprizone模型组比较,DHTS1处理明显减少cuprizone诱导的小鼠大脑胼胝体区髓鞘脱失,减少细胞凋亡,减少Iba-1^+阿米巴样小胶质细胞的面积,减少CD86^+细胞数目,增加CD163^+细胞数目。与CMC-Na联合LPS处理组相比,DHTS1明显减少CD16/32^+、TNF-α^+、iNOS^+小胶质细胞的百分比。结论DHTS1可以抑制cuprizone诱导的脱髓鞘和细胞凋亡,可能与DHTS1调节小胶质细胞极化,减轻中枢神经系统炎症反应有关。
Objective To evaluate whether dihydrotanshinoneⅠ(DHTS1)attenuates cuprizone-induced demyelination.Methods DHTS1 was dissolved in 5 g/L sodium carboxymethyl cellulose(CMC-Na).The cuprizone model was induced via feeding with the diet containing 2 g/L cuprizone.We administrated DHTS1 to the cuprizone-exposed mice.The mice were randomly divided into CMC-Na normal group,CMC-Na combined with cuprizone group and DHTS1 combined with cuprizone group.Myelin degeneration was checked by Luxol fast blue(LFB)staining and the immunohistochemical staining of myelin basic protein(MBP)and myelin proteolipid(PLP).Cell apoptosis was measured by TUNEL.Microglia polarization was evaluated by Iba-1,CD86 and CD163 immunohistochemical staining in vivo.The SIM-A9 cells cultured were divided into CMC-Na group,DHTS1 group,CMC-Na combined with LPS group and DHTS1 combined with LPS group.The expression of CD16/32,tumor necrosis factor-α(TNF-α),inducible nitric oxide synthase(iNOS)was analyzed by flow cytometry in vitro.Results Compared with CMC-Na combined with cuprizone group,DHTS1 treatment significantly attenuated myelin loss and cell apoptosis,reduced the area of Iba-1^+amoebic microglia and the number of CD86^+cells,while increased the number of CD163^+cells in the corpus callosum area of the brain.In addition,compared with CMC-Na combined with LPS group,DHTS1 obviously decreased the percentages of CD16/32^+,iNOS^+,TNF-α^+microglia.Conclusion DHTS1 can suppress cuprizone-induced demyelination and cell apoptosis through regulating the microglia polarization and mitigating inflammatory reaction in the central nerve system.
作者
徐芳
张晓娟
刘晓琴
张伟
佘晓岚
薛秀花
边疆
郭敏芳
尉杰忠
马存根
李艳花
XU Fang;ZHANG Xiaojuan;LIU Xiaoqin;ZHANG Wei;SHE Xiaolan;XUE Xiuhua;BIAN Jiang;GUO Minfang;YU Jiezhong;MA Cungen;LI Yanhua(Research Center of Neurobiology,Key Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine,Shanxi University of Chinese Medicine,Jinzhong 030619;Institute of Brain Science,Shanxi Key Laboratory of Inflammatory Neurodegenerative Diseases,Shanxi Datong University,Datong 037009;Ministry-of-Education Key Laboratory of Cell Proliferation and Regulation Biology,Beijing Normal University,Beijing 100875,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2020年第5期404-412,共9页
Chinese Journal of Cellular and Molecular Immunology
基金
大同市国际合作重点研发计划项目(2019123)
北京师范大学细胞增殖及调控生物学教育部重点实验室开放课题资助(LCPRB201803)
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(201805D111009-KF2019009)。
