摘要
目的探讨艾地苯醌是否通过抑制素2(prohibitin 2,PHB2)介导的线粒体自噬改善帕金森病(Parkinson disease,PD)模型小鼠行为学障碍。方法第一个小实验选取30只小鼠按照随机数字表法随机分为正常对照组、模型组和治疗组,每组10只,观测艾地苯醌对帕金森模型小鼠行为学的影响。第二个小实验选取20只小鼠按照随机数字法分为空白对照组、MPTP组、shRNA-PHB2组及shRNA-PHB2+MPTP组4组,每组5只,免疫荧光实验检测脑组织酪氨酸脱氢酶(TH)的改变。第三个小实验选取30只小鼠按照随机数字表法分为空白对照组、shRNA-PHB2组、MPTP+idebenone组、shRNA-PHB2+MPTP组、shRNA-PHB2+idebenone组和shRNA-PHB2+MPTP+idebenone组,每组5只,观测艾地苯醌对小鼠脑组织线粒体自噬的作用机制。将C57BL-6小鼠腹腔注射MPTP构建帕金森病动物模型,然后给予200 mg/kg艾地苯醌灌胃21 d,侧脑室显微注射腺相关病毒9(AAV9)shRNA-抑制素2(PHB2),抑制脑内PHB2的表达。Morris水迷宫实验检测小鼠行为学变化,免疫组化检测抑制PHB2对酪氨酸脱氢酶(TH)的改变,Western blot检测LC3、PHB2在小鼠中脑黑质中的蛋白表达。采用GraphPad 7.0和SPSS 22.0对数据进行统计分析。结果(1)第一个小实验中的水迷宫测试数据使用重复测量方差分析,测试1~7天小鼠逃避潜伏期组别-时间交互效应显著(F时间×组别=20.51,P<0.01);通过简单效应分析,与模型组相比,治疗组小鼠第5、6、7天逃避潜伏期明显缩短,差异有统计学意义(均P<0.05)。测试第7天,通过单因素方差分析比较,对照组与模型组,模型组与治疗组,对照组与治疗组之间均差异有统计学意义(t=-49.95,-21.81,28.14;均P<0.001)。第三个小实验中水迷宫测试数据通过重复测量方差分析显示,时间及组别交互作用显著(F时间×组别=42.11,P<0.01)。通过简单效应分析,与MPTP+idebenone组相比,shRNA-PHB2+MPTP+idebenone组潜伏期均明显延长(均P<0.05);与shRNA
Objective To investigate whether idebenone can improve behavioral disorders in mice with Parkinson disease(PD)by increasing PHB2 mediated mitophagy.Methods In the first small experiment,thirty mice were randomly divided into normal control group,model group and treatment group according to the random number table method,with 10 animals in each group.The aim of this study was to observe the effect of idebenone on the behavior of Parkinson disease model mice.In the second experiment,20 mice were randomly divided into blank control group,MPTP group,shRNA-PHB2 group and shRNA-PHB2+MPTP group,with 5 mice in each group.The changes of tyrosine dehydrogenase(TH)in brain tissue were detected by immunofluorescence assay.In the third experiment,30 mice were randomly divided into blank control group,shRNA-PHB2 group,MPTP+idebenone group,shRNA-PHB2+MPTP group,shRNA-PHB2+idebenone group and shRNA-PHB2+MPTP+idebenone group,with 5 animals in each group.The aim of this study was to investigate the effect of idebenone on mitochondrial autophagy in mouse brain.C57BL-6 mice were intraperitoneally injected with MPTP to establish the animal model of chronic PD.Then 200 mg/kg idebenone was given by gavage for 21 days.And the expression of PHB2 in brain was inhibited by microinjection of adeno-associated virus 9(AAV9)shRNA inhibin 2(PHB2)into lateral ventricle.The behavioral changes of the PD mice were detected by Morris water maze,and the changes of tyrosine dehydrogenase(TH)induced by inhibiting PHB2 were detected by immunohistochemistry.The protein expression of LC3 and PHB2 in substantia nigra of midbrain was detected by Western blot.The data were analyzed by GraphPad 7.0 and SPSS 22.0.Results(1)In the water maze test data of the first small experiment,the repeated measurement ANOVA showed that the group-time interaction effects of latency of mice from 1 to 7 days were significant(Ftime×group=20.51,P<0.05).Simple effect analysis showed that on the 5th,6th and 7th day,the incubation period of the treatment group was significantly sh
作者
闫俊强
刘安然
黄家瑞
吴剑男
马红霞
孙文杰
Yan Junqiang;Liu Anran;Huang Jiarui;Wu Jiannan;Ma Hongxia;Sun Wenjie(Depatment of Neurology,the First Affiliated Hospital,College of Clinical Medicine of Henan University of Science and Technology,Luoyang 471003,China;Neruomolecular Biology Laboratory,the First Affiliated Hospital,College of Clinical Medicine of Henan University of Science and Technology,Luoyang 471003,China)
出处
《中华行为医学与脑科学杂志》
CAS
CSCD
北大核心
2021年第1期15-21,共7页
Chinese Journal of Behavioral Medicine and Brain Science
基金
河南省医学科技攻关计划省部共建重点项目(SBGJ202002099)
河南省自然科学基金项目(18230041033)。
