MPTP诱导小鼠肝脏帕金森病样病理变化研究  

Study on MPTP-induced Parkinson's Disease-like Pathological Changes in Mice Liver

作　　者：韩奇文 陈颖 陈乃宏[1] 苑玉和[1] HAN Qiwen;CHEN Ying;CHEN Naihong;YUAN Yuhe(Institute of Materia Medica,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China)

机构地区：[1]中国医学科学院北京协和医学院药物研究所,北京100050

出　　处：《中国药物警戒》2021年第4期306-311,共6页Chinese Journal of Pharmacovigilance

基　　金：国家自然科学基金项目(81773925)。

摘　　要：目的研究1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)诱导的帕金森病(Parkinson's disease,PD)小鼠模型中肝脏生理功能变化和病理改变。方法将雄性C57BL/6小鼠随机分为空白对照组和模型组,皮下注射MPTP联合腹腔注射丙磺舒(MPTP/p),空白对照组以相同方法注射生理盐水,每4 d注射1次,共注射10次。苏木素-伊红染色(hematoxylin-eosin staining,HE)评估小鼠肝组织结构病理变化,免疫组化法观察肝脏组织不同形式α-突触核蛋白(α-synuclein,α-syn)的变化,以及8-羟基脱氧鸟苷(8-hydroxy-2 deoxyguanosine,8-OHdG)和4-羟基壬烯醛(4-hydroxy-2-nonenal,4-HNE)表达改变。通过使用自由氧(reactive oxygen species,ROS)荧光探针—二氢乙啶(dihydroethidium,DHE)染色检测肝脏部位的ROS水平。Western blot和免疫荧光检测肝组织内核转录相关因子2(Nuclear factor erythroid 2 related factor 2,Nrf2)在细胞核和细胞浆中的分布情况。结果MPTP/p联合诱导的PD小鼠模型中,肝组织表现出明显的炎症反应;硝基化形式的α-syn(n-syn)和寡聚体形式的α-syn(5G4)蛋白含量明显增加;肝组织内发生了脱氧核糖核酸(DNA)损伤及氧化应激反应;肝组织中的Nrf2总蛋白表达上调,胞浆中的Nrf2显著减少。结论PD致病相关因素能够诱导动物模型肝脏病理结构和生理功能发生显著变化,上述改变与氧化应激密切相关,α-syn可作为机体病变的指示分子。Objective To investigate the physiological function and pathological changes of the liver in a mouse model of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)induced Parkinson's disease(PD).Methods Male C57BL/6 mice were randomly divided into the control group and model group.The model group received subcutaneous injection of MPTP combined with an intraperitoneal injection of probenecid(MPTP/p)every 4 days with MPTP/p for 10 times,while the control group was injected with physiological saline in the same way.The pathological changes of the mouse liver were assessed by Hematoxylineosin staining(HE),while the changes of different forms ofα-synuclein(α-syn)in the liver,and those of expressions of 8-hydroxydeoxyguanosine(8-OHdG)and 4-hydroxy-2-Nonenal(4-HNE)were measured using an immunohistochemical method.The level of ROS in the liver was detected by dihydroethidium(DHE)staining with a fluorescent probe of reactive oxygen species(ROS).Western blot and immunofluorescence were used to detect the distribution of nuclear factor erythroid-2 related factor 2(Nrf2)in the nucleus and cytoplasm of liver tissue.Results In the PD mouse model induced by MPTP/p,liver tissue showed obvious inflammatory infiltration.The levels of the nitrated form ofα-syn(n-syn)and oligomeric form ofα-syn(5G4)protein increased.DNA damage and oxidative stress occurred in liver tissue.The expression of total Nrf2 protein in liver cells was up-regulated,but Nrf2 in the cytoplasm was reduced significantly.Conclusion PD pathogenic factors can induce significant changes in the pathological structure and physiological functions of the liver in model animals.The above changes are closely related to oxidative stress,andα-syn can be used as an indicator molecule for body disease.

关 键 词：1-甲基-4-苯基-1 2 3 6-四氢吡啶 帕金森病 肝脏 Α-突触核蛋白 氧化应激 

分 类 号：R994.1[医药卫生—毒理学]