松果菊苷抑制MPTP诱导的帕金森病模型小鼠神经胶质细胞激活及其机制研究  

Effect and Mechanism of Echinacoside on Inhibiting Glial Cell Activation in MPTP Mouse Model of Parkinson’s Disease

作　　者：张彦红[1] 刘美玲 朱磊 刘梦 姚丽伟 陈淑云 赵威[2] 王奇[2] ZHANG Yanhong;LIU Meiling;ZHU Lei;LIU Meng;YAO Liwei;CHEN Shuyun;ZHAO Wei;WANG Qi(The First People's Hospital of Guangzhou,Guangzhou 510180 Guangdong,China;Science and TechnologyInnovation Center,Guangzhou University of Chinese Medlicine,Guangzhou 510405Guangdong,China;GraduateSchool,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州市第一人民医院,广东广州510180 [2]广州中医药大学科创中心,广东广州510405 [3]广州中医药大学研究生院,广东广州510405

出　　处：《中药新药与临床药理》2021年第1期36-43,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省自然科学基金项目(2016A030313435)。

摘　　要：目的探讨松果菊苷对帕金森病(Parkinson’s disease,PD)模型小鼠脑部黑质致密部小胶质细胞活化与多巴胺(dopamine,DA)神经元损伤的作用,及其对多巴胺神经元-小胶质细胞生存微环境的影响。方法①松果菊苷药效实验:采用C57BL/6雄性小鼠,随机分为空白组、模型组、阳性药组(美满霉素,30 mg·kg^-1)、松果菊苷组(20 mg·kg^-1),除空白组、模型组注射等体积生理盐水外,其余组分别腹腔注射相应药物,每日1次,连续给药至取材前停止。给药3 d后,除空白组外,各组腹腔注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP,30 mg·kg^-1),每日1次,连续7 d,分别观察各组小鼠运动行为改变,脑部小胶质细胞特异性标志物(OX-42)和酪氨酸羟化酶(TH)阳性细胞数目;②松果菊苷药效机制研究:采用C57BL/6雄性小鼠,随机分为假手术组、模型组(IgG溶液,0.2μg·μL-1)、松果菊苷组(20 mg·kg^-1)、anti-Chemokine receptor组(AntiCX3CR1,0.2μg·μL-1),假手术组脑部注射等体积生理盐水,模型组脑部注射等体积IgG溶液,松果菊苷组腹腔注射松果菊苷溶液,Anti-CX3CR1组则在每次MPTP注射前1 h,侧脑室注射Anti-CX3CR1(5μL),造模方法同①,各组分别观察CX3CR1的表达水平的改变。结果松果菊苷腹腔注射给药后,可明显改善MPTP腹腔注射模型动物运动行为状态(P<0.05,P<0.01),增加TH的表达、降低OX-42免疫荧光表达强度(P<0.05,P<0.01),明显减轻模型小鼠的小胶质细胞激活及多巴胺能神经元的损伤。West Blot结果显示,Anti-CX3CR1组脑内CX3CR1表达水平与松果菊苷组表达一致。结论松果菊苷腹腔注射给药可较明显地降低MPTP造成的多巴胺能神经元损伤,其机制可能与抑制小胶质细胞CX3CR1激活有关。Objective To investigate the effects of echinacoside on the activation of microglia in substantia nigra compacta and the injury of DA neurons in Parkinson′s disease(PD) mice, and on the dopaminergic neuronmicroglia survival microenvironment.Methods①Pharmacodynamic experiment:Male C57 BL/6 mice were randomly divided into blank group, model group, positive drug group(minocycline, 30 mg·kg^-1)and echinacea glycoside group(20 mg·kg^-1). The blank group and the model group were injected with equal volume of normal saline,and the other groups were injected intraperitoneally with corresponding drugs,once a day for 3 d. The groups except blank group were intraperitoneally injected with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridin(MPTP,30 mg·kg^-1)for successive 7 days(once a day). The changes of motor behavior,complement receptor-3(OX-42)and the number of tyrosine hydroxylase(TH)-positive cells were observed. ② Pharmacodynamic mechanisms: Male C57 BL/6 mice were randomly divided into sham-operation group(normal saline,5 μL),model group(IgG solution,0.2 μg·μL-1),echinacea glycoside group(20 mg·kg^-1),and anti-chemokine receptor group(Anti-CX3 CR1,0.2 μg·μL-1). The injection volume of all groups was equal. IgG group and anti-chemokine receptor group were respectively injected IgG or anti-CX3 CR1 into the lateral ventricle. Animals in the anti-chemokine receptor group were injected with antiCX3 CR1 one hour before MPTP injection. Theexpression of CX3 CR1 was investigated.ResultsAfter intraperitoneal injection of echinacoside for 10 days,the behaviors of MPTP-treated mice were altered(P<0.05,P<0.01),the immunofluorescence expression of OX-42 and TH were all restored(P<0.05,P<0.01),and the level of CX3 CR1 was decreased.ConclusionIntra-peritoneal injection of echinacoside can significantly reduce the damage of dopaminergic neurons caused by MPTP, which may be related with the inhibition of echinacoside on microglia activation.

关 键 词：帕金森病 松果菊苷 小胶质细胞 多巴胺神经元 CX3CR1 小鼠 

分 类 号：R285.5[医药卫生—中药学]