丁苯酞联合美多芭对帕金森病伴抑郁模型小鼠黑质纹状体Ⅱ型髓系细胞触发受体及小胶质细胞炎症标志物表达的影响  

Effects of butylphthalide combined with medoba on expression of TREM2 and microglia inflammatory markers in substantia nigra striatum of Parkinson’s disease with depression model mice

作　　者：刘斌 杨浩辉 任伯 范少凯 毛文静 王雅楠 吴小坤 LIU Bin;YANG Haohui;REN Bo;FAN Shaokai;MAO Wenjing;WANG Yanan;WU Xiaokun(First Department of Neurology,the Affiliated Hospital of North China University of Science and Technology,Tangshan 063000,China)

机构地区：[1]华北理工大学附属医院神经内一科,河北唐山063000

出　　处：《陕西医学杂志》2022年第12期1477-1482,共6页Shaanxi Medical Journal

基　　金：河北省政府资助临床医学优秀人才培养项目(2019046)。

摘　　要：目的:探讨丁苯酞联合美多芭对帕金森病伴抑郁模型小鼠黑质纹状体Ⅱ型髓系细胞触发受体(TREM2)及小胶质细胞M1型炎症标志物[白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)]及M2型炎症标志物[IL-10和精氨酸酶-1(Arg-1)]表达的影响。方法:随机将40只C57BL/6成年雄性小鼠分正常对照组(对照组)、帕金森病伴抑郁模型组(模型组)、美多芭治疗组和丁苯酞联合美多芭治疗组,每组10只。腹腔注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP,30 mg/kg×7 d)制备帕金森病小鼠模型。建立帕金森病模型后,以慢性不可预知温和应激诱导产生帕金森病伴抑郁小鼠模型。美多芭治疗组给予美多芭6.25 mg/kg灌胃治疗,1次/d,持续7 d。丁苯酞联合美多芭治疗组给予丁苯酞120 mg/kg和美多芭6.25 mg/kg灌胃治疗,1次/d,持续7 d。采用免疫组化定性分析黑质纹状体内α-突触核蛋白的阳性表达情况。采用酶联免疫吸附(ELISA)法测定5-羟色胺(5-HT)含量。采用蛋白免疫印迹法检测TREM2蛋白表达水平。采用蛋白免疫印迹和PCR检测小胶质细胞炎症标志物IL-1β、TNF-α、IL-10、Arg-1蛋白及mRNA表达水平。结果:模型组小鼠黑质纹状体部位可见大量α-突触核蛋白异常积聚;美多芭治疗组及丁苯酞联合美多芭治疗组α-突触核蛋白阳性表达明显减少,染色变浅,以丁苯酞联合美多芭治疗组减少更明显。与对照组比较,模型组5-HT含量明显减低(P<0.05);与模型组比较,美多芭治疗组及丁苯酞联合美多芭治疗组5-HT含量明显增加,以丁苯酞联合美多芭治疗组增加更明显(均P<0.05)。与对照组比较,模型组TREM2明显减低(P<0.05);与模型组比较,美多芭治疗组及丁苯酞联合美多芭治疗组TREM2明显增加,以丁苯酞联合美多芭治疗组增加更明显(均P<0.05)。与对照组比较,模型组小胶质细胞M1型炎症标志物IL-1β和TNF-α明显升高,M2型标炎症标志物IL-10和Arg-1明显下降(均Objective:To explore the effects of butylphthalide combined with medoba on TREM2 and microglia M1 markeRIL-1β,TNF-αand M2 markeRIL-10,Arg-1 in substantia nigra striatum of Parkinson’s disease with depression model mice.Methods:Forty C57BL/6 adult male mice were randomly divided into normal control group(control group),Parkinson’s disease with depression model group(model group),medoba treatment group and butylphthalide combined with medoba treatment group,with 10 mice in each group.The mouse model of Parkinson’s disease was prepared by intraperitoneal injection of MPTP(30 mg/kg×7 d).AfteRthe establishment of Parkinson’s disease model,the mouse model of Parkinson’s disease with depression was induced by chronic unpredictable mild stress.The medoba treatment group was given medoba 6.25 mg/kg by gavage once a day foR7 days.The butylphthalide combined with medoba treatment group was given medoba 6.25 mg/kg and butylphthalide 120 mg/kg by gavage once a day foR7 days.The positive expression ofα-synuclein in substantia nigra striatum was qualitatively analyzed by immunohistochemistry,and the content of serotonin was determined by enzyme-linked immunosorbent assay(ELISA),and the level of TREM2 protein in substantia nigra striatum was detected by Western blot,and microglia inflammatory markers IL-1β,TNF-α,IL-10,Arg-1 level changes of microglia were detected by Western blot and PCR.Results:A large numbeRofα-synuclein abnormal accumulation in the model group were found in the substantia nigra and striatum,the positive expression ofα-synuclein decreased significantly and the staining became lighteRin medoba treatment group and butylphthalide combined with medoba treatment group,especially in butylphthalide combined with medoba treatment group.In comparison with the control group,5-HT content in the model group decreased significantly(P<0.05);in comparison with the model group,5-HT content increased significantly in medoba treatment group and butylphthalide combined with medoba treatment group,especially in

关 键 词：帕金森病 抑郁 丁苯酞 美多芭 Ⅱ型髓系细胞触发受体 小胶质细胞 

分 类 号：R742.5[医药卫生—神经病学与精神病学]