铝对斑马鱼幼鱼运动行为的影响及其机制  

Effect of aluminum on locomotor activity of zebrafish larvae and its mechanism

作　　者：谈勇 许梦川 王远 张芳 李小满 陈豪 赵宝全 汪雪雁[1] TAN Yong;XU Meng-chuan;WANG Yuan;ZHANG Fang;LI Xiao-man;CHEN Hao;ZHAO Bao-quan;WANG Xue-yan(College of Tea and Food Science ＆ Technology,Anhui Agricultural University,Hefei 230036,China;State Key Laboratory of Toxicology and Medical Countermeasures,Institute of Toxicology and Pharmacology,Academy of Military Medical Sciences,Academy of Mililtary Sciences,Beijing 100850,China;State Key Laboratory of Kidney Disease,Chinese PLA General Hospital,Beijing 100853,China)

机构地区：[1]安徽农业大学茶与食品科技学院,安徽合肥230036 [2]军事科学院军事医学研究院毒物药物研究所抗毒药物与毒理学国家重点实验室,北京100850 [3]解放军总医院肾脏疾病国家重点实验室,北京100853

出　　处：《中国药理学与毒理学杂志》2018年第3期208-214,共7页Chinese Journal of Pharmacology and Toxicology

摘　　要：目的研究铝对斑马鱼幼鱼运动行为的影响及其分子机制。方法将受精后3 d的斑马鱼幼鱼随机分别暴露于AlCl_3(0,50,100和150μg·L-1)48,72或96 h,利用Noldus斑马鱼幼鱼行为学检测系统检测其行为,HE染色检测眼的病理变化,生化法检测活性氧(ROS)水平,荧光定量PCR检测血脑屏障及神经系统相关基因m RNA表达水平。结果与正常对照组相比,AlCl_3暴露组斑马鱼幼鱼运动轨迹杂乱无章,部分个体停止活动;暴露48 h,AlCl_3150μg·L-1组运动距离和速度显著下降(P<0.01);暴露72和96 h,各浓度组运动距离和速度均显著下降(P<0.01)。病理检测结果表明,各组斑马鱼幼鱼无明显的眼组织病理改变。与正常对照组相比,暴露48 h,AlCl_3100和150μg·L-1组斑马鱼幼鱼体内ROS水平显著升高(P<0.01);暴露72和96 h,AlCl_3各浓度组ROS水平均显著升高(P<0.01)。荧光定量PCR检测结果显示,与正常对照组相比,AlCl_3暴露48 h,50μg·L-1组淀粉样前体蛋白b(appb)m RNA表达明显升高(P<0.01),100μg·L-1组早老素1(psen1)和胶质细胞原纤维酸性蛋白(gfap)m RNA表达显著降低(P<0.01,P<0.05),150μg·L-1组psen1,生长相关蛋白43(gap43)和gfap m RNA表达均显著降低(P<0.01);AlCl_3暴露72 h,50μg·L-1组gap43和gfap m RNA表达明显降低(P<0.01),100和150μg·L-1组appb和psen1 m RNA表达显著升高(P<0.01),gap43和gfap m RNA表达明显降低(P<0.01);暴露96 h,AlCl_350μg·L-1组appb和gap43 m RNA表达显著升高(P<0.01),100μg·L-1组appb和psen1 m RNA表达显著升高(P<0.01),gap43和gfap m RNA表达明显降低(P<0.01,P<0.05),150μg·L-1组appb和psen1 m RNA表达显著升高(P<0.01),密封蛋白(cldn5a),gap43和gfap m RNA表达明显降低(P<0.01,P<0.05)。结论铝暴露可导致斑马鱼幼鱼运动行为发生变化,其机制可能与体内ROS水平升高,以及cldn5a,appb,psen1,gap43和gfap基因表达的变化有关。OBJECTIVE To explore the effect of aluminum on locomotor activity of zebrafish larvae and its possible mechanism. METHODS Zebrafish larvae（3-day post fertilization） were exposed to different concentrations of AlCl3（0, 50, 100 and 150 μg·L^-1） for 48, 72 or 96 h. The locomotor activity of zebrafish larvae was recorded by Noldus Tracking System, the morphological changes in zebrafish larvae eyes were observed by HE staining, the level of reactive oxygen species（ROS） was determined by biochemical method, while the mRNA expression level of genes related to the blood-brain barrier and nervous system was analyzed by quantitative polymerase chain reaction（q PCR）. RESULTS Compared with normal control group, the movement trajectories of zebrafish larvae exposed to AlCl3 were disorganized, and even some stopped swimming. After exposure for 48 h, the movement distance and speed were significantly decreased in AlCl3150 μg·L^-1 group（P〈0.01）. After exposure for 72 and 96 h, the movement distance and speed were significantly reduced in all concentrations of AlCl3（P〈0.01）. Pathological examination showed that there were no obvious histopathological changes in the eyes of zebrafish exposed to AlCl3. The ROS level of zebrafish larvae was increased in AlCl3100 and 150 μg·L^-1 groups after 48 h exposure（P〈0.05, P〈0.01） and in all concentrations of AlCl3 group after 72 and 96 h exposure（P〈0.01）. The q PCR results showed that compared with control group, after AlCl3 exposure of 48 h,the mRNA expression of amyloid precursor protein b（appb） was significantly increased（P〈0.01） in 50 μg·L^-1 group, the mRNA expressions of presenilin 1（psen1） and glial fibrillary acidic protein（gfap） were downregulated（P〈0.01, P〈0.05） in 100 μg·L^-1 group, and the mRNA expressions of psen1, growth-associated protein 43（gap43） and gfap were reduced in 150 μg·L^-1 group（P〈0.01）. After AlCl3 exposure for 72 h,the mRNA expressions of gap43 and gfap were decreased signifi

关 键 词：铝 斑马鱼 神经毒性 

分 类 号：R99[医药卫生—毒理学]