雷帕霉素和3-甲基腺嘌呤对帕金森病小鼠运动行为及黑质自噬相关蛋白LC3的影响  

Effects of rapamycin and 3-MA on motor behavior and autophagy related protein LC3 in PD model mice

作　　者：吴忧[1] 张荣博[1] 粱顺利 朱悦红 徐彬[1] Wu You Zhang Rongbo Liang Shunli Zhu Yuehong Xu Bin(Department of Neurology, the Second Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310005,China)

机构地区：[1]浙江中医药大学附属第二医院神经内科,杭州310005

出　　处：《中华行为医学与脑科学杂志》2016年第11期961-965,共5页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：浙江省医药卫生科技计划项目（201471632）;浙江省教育厅项目（Y201431457）;浙江省中医药科技计划项目（青年人才基金项目）（2015ZQ017）

摘　　要：目的探讨自噬诱导剂雷帕霉素和自噬抑制剂3-甲基腺嘌呤（3-MA）对C57BL／6帕金森病（PD）模型小鼠运动行为及自噬相关蛋白LC3的影响。方法取40只C57BL／6雄性小鼠随机分为实验组和正常对照组，实验组又分为MPTP模型组、雷帕霉素组、3-MA组，每组10只。实验组采用MPTP腹腔注射建立PD模型，正常对照组给予等量生理盐水腹腔注射。雷帕霉素组和3-MA组在MPTP注射的同时分别予雷帕霉素、3-MA腹腔注射。各组小鼠在最后1次MPTP或生理盐水注射后1d、7d、14d分别进行行为学测试，最后1次行为学测试后处死小鼠取中脑黑质脑组织，WesternBlot检测自噬相关蛋白LC3的表达。结果与MPTP模型组比较，雷帕霉素组小鼠的爬杆时间明显缩短[（14．89±1．14）s vs（16．24±1．39）s，P〈0．05；（15．18±1．36）s vs（17．93±1．11）s，P〈0．01]，悬挂分值增加[（1．7±0．5）分粥（1．2±0．4）分，P〈0．05；（1．5±0．5）分vs（1．1±0．3）分，P〈0．05]，旷场实验的总距离[（5875．3±1148．9）cm vs（5061．5±773．1）cm，P〈0．05；（5753．2±1106．7）cm vs（4669．3±969．1）cm，P〈0．01]及平均速度[（19．29±2．35）cm／s vs（16．47±3．01）cm／s，P〈0．05；（18．71±2．71）cm／s vs（15．80±2．50）cm／s，P〈0．01]均增加，LC3-Ⅱ蛋白表达增加，而3-MA组小鼠的爬杆时间延长[（19．92±1．61）s vs（17．93±1．11）s，P〈0．05]，旷场实验总距离[（3879．7±575．0）cm vs（4669．3±969．1）cm），P〈0．05]及平均速度[（13．34±1．87）cm／s vs（15．80±2．50cm／s，P〈0．05]减少，LC3-Ⅱ蛋白表达减少。结论自噬诱导剂雷帕霉素可改善PD小鼠的行为学症状，增加黑质自噬相关蛋白LC3的表达，而自噬抑制剂3-MA则会加重PD小鼠的行为学障碍，减少黑质LC3的表达．调节自噬的药物可能是PD防治的新方向。Objective To investigate the effects of rapamyein which is an autophagy inducer and 3- adenine （3-MA） which is an autophagy inhibitor on motor behavior and autophagy related protein LC3 in C57BL/6 mice of Parkinson＇s disease（PD）. Methods 40 C57BL/6 male mice were randomly divided into control group and experimental groups which consist of MPTP model group, rapamyein group and 3-MA group, with 10 in each group. Mice in experimental groups received intraperitoneal injection with MPTP to es- tablish PD models, and mice in control group received intraperitoneal injection with the same amount of saline solution for 1 week. Mice in rapamyein group received intraperitoneal injection with rapamyein and mice in 3- MA group received intraperitoneal injection with 3-MA at the same time when MPTP was injected.Animal be- havior detections were carried out on the lth,7th and 14th day after the last injection.After the last injection, mice were sacrificed and sections of midbrain nigra were gained for the detection of expression of autophagy specificity protein LC3 by Western Blot. Results Compared with MPTP model group, rapamycin could im- prove the general eondition and behavioral manifestation of mice in pole test （ （ 14.89 ± 1.14 ） s vs （ 16.24 ±1.39）s, P〈0.05; （ 15.18±1.36）s vs（17.93±1.11s） , P〈0.01） ,traction test（ （1.7±0.5） vs （ 1.2±0.4）, P〈 0.05; （1.5±0.5） vs （ 1.1 ±0.3 ）, P〈0.05）and open field test which contained total activity distance（ （5 875.3 ±1148.9）cm vs （5 061.5±773.1）cm, P〈0.05; （5 753.2±1 106.7）cm vs （4 669.3±969.1）cm, P〈0.01） and average speed（ （19.29±2.35） cm/s vs （ 16.47±3.01 ） cm/s, P〈0.05 ; （ 18.71 ±2.71 ） cm/s vs （15.80±2.50） cm/s, P〈0.01 ） ,while 3-MA aggravated behavioral deficits of mice in pole test（ 19.92± 1.61s vs 17.93± 1.11 s, P〈O.05）, and both total activity distance （ （ 3 879.7±575.0） cm vs （4 669.3±969.1 ） cm, P〈0.05） and aver- age speed（ （ 13.3

关 键 词：雷帕霉素 3-甲基腺嘌呤 帕金森病 LC3 自噬 

分 类 号：R742.5[医药卫生—神经病学与精神病学]