姜黄素干预通过促进自噬改善氯化锰所致的大鼠神经行为损伤  

Curcumin alleviates the manganese-induced neurotoxicity by promoting autophagy in rat models of manganism

作　　者：来丽叶 窦长松 智翠娜 陈洁 马雪 赵鹏[1] 姚碧云[1] LAI Li-ye;DOU Chang-song;ZHI Cui-na;CHEN Jie;MA Xue;ZHAO Peng;YAO Bi-yun(Department of Toxicology, Peking University School of Public Health, Beijing Key Laboratory of Toxicological Research and Risk Assessment for Food Safety, Beijing 100191, China)

机构地区：[1]北京大学公共卫生学院毒理学系,食品安全毒理学研究与评价北京市重点实验室,北京100191

出　　处：《北京大学学报（医学版）》2022年第3期400-411,共12页Journal of Peking University:Health Sciences

基　　金：北京市自然科学基金(7192099、7150284)。

摘　　要：目的:探讨姜黄素(curcumin,CUR)对锰中毒大鼠神经损伤的干预效应及其机制。方法:成年雄性SD大鼠60只,随机分为5组,每组12只。(1)空白对照组:通过腹腔注射(intraperitoneal injection,ip)给予0.9%(质量分数)生理盐水,并通过灌胃(intragastric,ig)给予双蒸水(double distilled water,ddH_(2)O);(2)锰中毒模型组:ip给予MnCl_(2)15 mg/kg(Mn^(2+) 6.48 mg/kg),ig给予ddH_(2)O;(3)单独姜黄素组:ip给予0.9%生理盐水,ig给予CUR 300 mg/kg;(4)MnCl_(2)+CUR1组:ip给予MnCl_(2)15 mg/kg,ig给予CUR 100 mg/kg;(5)MnCl_(2)+CUR2组:ip给予MnCl_(2)15 mg/kg,ig给予CUR 300 mg/kg。5组大鼠每周给药5 d,连续给予4周。旷场实验、转棒实验检测动物的探索行为、焦虑抑郁状态、运动及平衡能力,Morris水迷宫(Morris water maze,MWM)实验检测动物的学习、记忆能力。每组6只大鼠进行脑纹状体HE染色和免疫组织化学(immunohistochemistry,IHC)检查,并且其中2只大鼠一侧纹状体进行透射电镜(transmission electron microscope,TEM)观察。每组另外6只大鼠的一侧纹状体利用电感耦合等离子体质谱(ICP-MS)测定锰含量,另一侧纹状体用免疫印迹(Western blotting,WB)检测α-突触核蛋白(α-Synuclein,α-Syn)、细胞质和细胞核转录因子EB(transcription factor EB,TFEB)、雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)、p-mTOR、Beclin1、P62和微管相关蛋白轻链蛋白3(microtubule-associated protein light chain-3,LC3)表达水平,脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(terminal deoxynucleotidyl transterase-mediated dUTP nick end labeling;TUNEL)检测大鼠纹状体多巴胺能神经元凋亡。结果:旷场和转棒实验中,锰中毒模型组水平运动距离、直立次数、中央区运动时间和运动距离、在棒时间、在棒圈数较空白对照组显著降低(P<0.05),而MnCl_(2)+CUR2组的上述指标均较锰中毒模型组显著增加(P<0.05)。MWM定位航行实验第3、4天时,锰中毒模型组逃避潜伏期和�Objective:To investigate the protective effects of curcumin(CUR)and its mechanism on a rat model of neurotoxicity induced by manganese chloride(MnCl_(2)),which mimics mangnism.Methods:Sixty male SD rats were randomly divided into 5 groups,with 12 rats in each group.Control group received 0.9%saline solution intraperitoneally(ip)plus double distilled water(dd)H_(2)O intragastrically(ig),MnCl_(2) group received 15 mg/kg MnCl_(2)(Mn^(2+) 6.48 mg/kg)intraperitoneally plus dd H_(2)O intragastrically,CUR group received 0.9%saline solution intraperitoneally plus 300 mg/kg CUR intragastrically,MnCl_(2)+CUR1 group received 15 mg/kg MnCl_(2) intraperitoneally plus 100 mg/kg curcumin intragastrically,MnCl_(2)+CUR2 group received 15 mg/kg MnCl_(2) intraperitoneally plus 300 mg/kg CUR intragastrically,5 days/week,4 weeks.Open-field and rotarod tests were used to detect animals’exploratory behavior,anxiety,depression,movement and balance ability.Morris water maze(MWM)experiment was used to detect animals’learning and memory ability.ICP-MS was used to investigate the Mn contents in striata.The rats per group were perfused in situ,their brains striata were removed by brains model and fixed for transmission electron microscope(TEM),histopathological and immunohistochemistry(ICH)analyses.The other 6 rats per group were sacrificed.Their brains striata were removed and protein expression levels of transcription factor EB(TFEB),mammalian target of rapamycin(mTOR),p-mTOR,Beclin,P62,microtubule-associated protein light chain-3(LC3)were detected by Western blotting.Terminal deoxynucleotidyl transterase-mediated dUTP nick end labeling(TUNEL)staining was used to determine neurocyte apoptosis of rat striatum.Results:After exposure to MnCl_(2) for four weeks,MnCl_(2)-treated rats showed depressive-like behavior in open-field test,the impairments of movement coordination and balance in rotarod test and the diminishment of spatial learning and memory in MWM(P<0.05).The striatal TH+ neurocyte significantly decreased,eosinophilic cells,aggr

关 键 词：锰中毒 姜黄素 转录因子EB 自噬 

分 类 号：R135.1[医药卫生—劳动卫生]