邻苯二甲酸二异丁酯对小鼠海马cAMP/PKA-CREB信号转导通路的影响  

Effects of DiBP on the cAMP/PKA-CREB-BDNF signaling pathway of hippocampus in mice

作　　者：马宁 王小丹 高芃 徐海滨 

机构地区：[1]国家食品安全风险评估中心,北京100021

出　　处：《卫生研究》2013年第3期405-409,共5页Journal of Hygiene Research

基　　金：国家科技支撑计划课题(No.2006BAK02A07)

摘　　要：目的观察邻苯二甲酸二异丁酯(DiBP)对小鼠海马cAMP/PKA-CREB信号通路的影响。方法 30只雄性昆明小鼠适应性喂养3天后,按体重随机分为一个对照组和4个实验组(Ⅰ、Ⅱ、Ⅲ、Ⅳ)。对照组小鼠灌胃给予玉米油溶剂;实验组小鼠灌胃给予含DiBP的玉米油溶液,剂量分别为:50、250、500、1000mg/kg BW。实验周期8周,普通饲料喂饲,自由饮水。实验结束后冰浴条件下取海马,采用酶联免疫法测定环磷酸腺苷(cAMP)含量;蛋白印迹法测定磷酸化蛋白激酶A(P-PKA C)、磷酸化环磷腺苷反应元件结合蛋白(P-CREB)表达水平;实时荧光定量PCR法测定CREB、脑源性神经营养因子(BDNF)、c-fos和c-jun的mRNA表达水平。结果第Ⅳ组海马组织中cAMP含量和P-PKA C蛋白表达水平明显低于对照组,差异具有统计学意义(P<0.05);各剂量组P-CREB蛋白表达量均低于对照组,其中第Ⅱ组与对照组相比差异有统计学意义(P<0.05),同时CREB、BDNF、c-fos和c-jun的mRNA表达水平与对照组相比都呈不同程度的下调。结论小鼠海马cAMP/PKA-CREB信号通路中多种信号分子的表达异常,可能是DiBP导致小鼠认知损伤的重要机制之一。Objective To explore the effects of diisobutyl phthalate（DiBP） on the cAMP/PKA-CREB signaling pathway of hippocampus in mice.Methods Accommodating 30 KunMing mice to the animal house for 3 days,then dividing the mice into 5 groups according to their weights.That is,one control group and four experimental groups（Ⅰgroup,50mg/kg BW;Ⅱ group,250mg/kg BW;Ⅲ group,500mg/kg BW;Ⅳ group,1000mg/kg BW）.The mice were fed with the corn oil in control group,and the other groups were fed with the related dose of DiBP mixture by gavages last for 8 weeks.At the end of experimental time,the mice were killed,and the tissue samples of hippocampus were taken immediately.The content of cAMP was determined by ELISA,and p-CREB,P-PKA C were measured by western blotting,while mRNA expressions including CREB,BDNF,c-fos and c-jun were checked by RT-PCR.Results The cAMP content and the p-PKA C protein of hippocampus in Ⅳ group was significantly less than control group（P0.05） and compared with control group,the p-CREB protein of hippocampus in Ⅱ group decreased（P0.05）,while the relative level of CREB,BDNF,c-fos and c-jun mRNA were down-regulated in all experimental groups.Conclusions The abnormal changes of expression of the signal molecules composing the cAMP/PKA-CREB signaling pathway were observed in the hippocampus of DiBP exposure mice.This might be one of the possible mechanisms of DiBP induced cognitive impairment.

关 键 词：邻苯二甲酸二异丁酯 海马 环磷腺苷反应元件结合蛋白 

分 类 号：R338.64[医药卫生—人体生理学] R994.6[医药卫生—基础医学]