天麻活性成分20C对6-羟基多巴胺诱导的PC12细胞损伤及帕金森病模型小鼠的保护作用  

Protective effect of 20C,an active ingredient of Gastrodiae Rhizoma,against 6-hydroxydopamine-induced PC12 cell damage and Parkinson disease model mice

作　　者：杜雨生 张钊[2] 何新[1,3] 陈乃宏 DU Yu-sheng;ZHANG Zhao;HE Xin;CHEN Nai-hong(School of Chinese Materia Medica,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;Institute of Materia Medica,Chinese Acadency of Medical Sciences&Peking Union Medical College,Beijing 100050,China;School of Chinese Materia Medica,Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]天津中医药大学中药学院,天津301617 [2]中国医学科学院北京协和医学院药物研究所,北京100050 [3]广东药科大学中药学院,广东广州510006

出　　处：《中国药理学与毒理学杂志》2021年第12期897-903,共7页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金(81603316);中国医学科学院医学与健康科技创新工程经费资助(2016-I2M-1-004)。

摘　　要：目的研究天麻活性成分20C对6-羟基多巴胺(6-OHDA)诱导的PC12细胞损伤及帕金森病(PD)模型小鼠的保护作用。方法(1)用6-OHDA 100μmol·L^(-1)单独(模型组)或与20C 0.01,0.10和1.00μmol·L-1共同(模型+20C组)孵育PC12细胞6 h,采用MTT法检测细胞存活率;用6-OHDA 100μmol·L^(-1)和20C1.00μmol·L^(-1)共同孵育PC12细胞6 h,采用免疫荧光染色法检测细胞DNA和RNA氧化损伤。(2)C57BL/6小鼠分为正常对照组、模型组、模型+左旋多巴(L-dopa)组和模型+20C组。采用单侧纹状体双位点注射6-OHDA方法制备PD小鼠模型,造模3周后,模型+L-dopa组和模型+20C组小鼠开始ig给予20C 100 mg·L^(-1)或L-dopa 40 mg·g^(-1),连续给药4周后,采用免疫组化法观察小鼠纹状体和黑质酪氨酸羟化酶(TH)的表达;Western印迹法检测黑质TH表达水平;透射电镜观察小鼠黑质神经元超微结构。结果(1)与细胞对照组比较,模型组PC12细胞存活率显著下降(P<0.01),细胞DNA/RNA氧化损伤明显增加(P<0.01);与模型组比较,模型+20C 0.10和1.00μmol·L^(-1)组细胞存活率显著升高(P<0.05,P<0.01),模型+20C 1.00μmol·L^(-1)组细胞DNA和RNA氧化损伤明显降低(P<0.05)。(2)与正常对照组相比,模型组小鼠纹状体和黑质区TH阳性染色区域减少,黑质区TH蛋白表达水平显著减少(P<0.01),黑质区神经元核膜皱缩,电子密度增加,细胞周围肿胀;与模型组相比,模型+20C 100 mg·kg^(-1)组纹状体和黑质区TH阳性染色区域增加,黑质区TH蛋白表达显著增加(P<0.05),神经元核膜皱缩及细胞周围肿胀程度减轻。结论20C能够降低6-OHDA诱导的PC12细胞死亡率,并减轻6-OHDA诱导的PD模型小鼠黑质和纹状体病理损伤。OBJECTIVE To study the protective effect of 20 C,an active ingredient of Gastrodiae Rhizoma,against 6-hydroxydopamine(6-OHDA)-induced PC12 cell damage and Parkinson disease(PD) model mice.METHODS PC12 cells were treated with 6-OHDA 100 μmol·L^(-1) and simultaneously given with 20 C 0.01,0.10 and 1.00 μmol·L^(-1) for 6 h,before the cell survival rate was detected by MTT assay.PC12 cells were treated using 6-OHDA 100 μmol·L^(-1) and simultaneously given 20 C 1.00 μmol·L^(-1) for 6 h,before the DNA and RNA oxidative damage was detected by cell immunofluorescence staining assay.C57 BL/6 mice were divided into the normal control,model,model+levodopa(L-dopa) and model+20 C groups.A mice model of PD was established by 6-OHDA injection at two sites in the unilateral striatum.Three weeks after 6-OHDA injection,mice in model+L-dopa and model+20 C groups were ig given 20 C100 mg·kg^(-1) or L-dopa 40 mg·kg^(-1) for 4 weeks.The tyrosine hydroxylase(TH) positive areas of the striatum and substantia nigra were detected by immunohistochemical staining assay.The TH protein expression in the substantia nigra was detected by Western blotting.Ultrastructural damage of neurons in the substantia nigra was detected by transmission electron microscopy.RESULTS Compared with the cell control group,the survival rate was decreased significantly(P<0.01) and the mean fluorescence intensity was significantly enhanced(P<0.01) in the model group.Compared with the model group,the survival rates in the model+20 C 0.10 μmol·L^(-1) and 1.00 μmol·L^(-1) group were significantly improved(P<0.05,P<0.01) and the mean fluorescence intensity in the model+20 C 1.00 μmol·L^(-1) group was significantly reduced(P<0.05).Compared with the normal control group,TH positive areas in the striatum and substantia nigra were decreased,TH protein expression in the substantia nigra was significantly decreased(P<0.01),the nuclear membrane of neurons in the substantia nigra was shrunken,electron density was increased and the surrounding cells were swollen

关 键 词：天麻 20C 帕金森病 6-羟基多巴胺 

分 类 号：R964[医药卫生—药理学] Q244[医药卫生—药学]