钙稳态在DBP诱导胚胎大鼠皮层神经元凋亡中的作用  

Role of Calcium Homeostasis in Apoptosis of Embryonic Rat Cortical Neurons Induced by Dibutylphthalate

作　　者：王文娟[1] 杨光红[1] 曾奇兵[1] 喻馨兰[1] 汪文云[1] 

机构地区：[1]贵州医科大学公共卫生学院职业卫生与职业医学教研室,贵州贵阳550025

出　　处：《环境与职业医学》2016年第3期228-232,共5页Journal of Environmental and Occupational Medicine

基　　金：贵阳市科技计划项目(编号:筑科合同[2013103]19号;贵州省科技计划(编号:黔科合LH字[2014]7102)

摘　　要：[目的]研究钙稳态在邻苯二甲酸二丁酯(DBP)诱导大鼠皮层神经元凋亡中的作用。[方法]取胚胎大鼠皮层神经元,用25、50、100 mg/L的DBP进行细胞染毒,同时用加入10μmol/L钙络合剂(BAPTA-AM)和上述各浓度的DBP进行细胞培养,检测各组细胞的凋亡率、细胞内钙浓度(Fluo-3/AM荧光强度)、活性氧浓度以及线粒体膜电位。[结果]25、50、100 mg/L DBP染毒组的神经元凋亡率[(12.27±0.93)%、(18.53±0.87)%、(27.63±1.06)%],荧光强度[(10 623±826)%、(14 815±732)%、(18 328±708)%],细胞内活性氧的发生率[(10.21±1.05)%、(29.23±1.35)%、(34.68±1.03)%]及神经元线粒体膜电位下降率[(30.71±1.81)%、(45.30±3.25)%、(53.00±4.29)%]均高于对照组,差异均有统计学意义(P<0.05)。随着DBP染毒剂量的增加,上述各项指标均呈剂量-效应关系(r=0.977,r=0.970,r=0.962,r=0.914,均P<0.05);BAPTA-AM干预后,与DBP同剂量组比较皮层神经元凋亡率有所降低,荧光强度明显下降,线粒体膜电位下降率降低,细胞内活性氧的发生率降低(均P<0.05)。[结论]DBP可诱导大鼠皮层神经元凋亡,钙络合剂通过降低钙离子浓度,缓解DBP诱导的皮神经元的凋亡,表明钙稳态失衡促进DBP诱导的大鼠皮层神经元凋亡。[Objective] To study the role of calcium homeostasis in the apoptosis of rat cortical neurons induced by dibutylphthalate（DBP）.[Methods] Neurons were sampled from the cortex of pregnant rats.The cortical neurons were treated with 25,50,and 100 mg/L DBP;meanwhile,10 μmol/L calcium chelator（BAPTA-AM） was added into the above combinations.The apoptosis rates,intracellular free calcium concentrations [Ca2＋]i,reactive oxygen species（ROS）,and mitochondrial membrane potential of the cortical neurons were detected by flow cytometry.[Results] After treated with 25,50,and 100 mg/L DBP,the apoptosis rate [（12.27±0.93）%,（18.53±0.87）%,and（27.63±1.06）%,respectively],Fluo-3/AM fluorescence intensity [（10 623±826）%,（14 815±732）%,and（18 328±708）%],level of ROS [（10.21±1.05）%,（29.23±1.35）%,and（34.68±1.03）%],and decrease rate of mitochondrial membrane potential [（30.71±1.81）%,（45.30±3.25）%,and（53.00±4.29）%] were all higher than those of the control group（all Ps 〈 0.05） in a dose-response manner（r=0.977,r=0.970,r=0.962,r=0.914,all Ps 〈 0.05）.After treated with BAPTAAM,the apoptosis rates,Fluo-3/AM fluorescence intensity,level of ROS,and decrease rate of mitochondrial membrane potential of the cortical neurons were decreased compared with corresponding DBP only groups（all Ps 〈 0.05）.[Conclusion] DBP could induce rat cortical neuron apoptosis,and BAPTA-AM could blunt the apoptosis through decreasing [Ca2＋]i,suggesting that the disequilibrium of calcium homeostasis might promote the apoptosis of rat cortical neurons induced by BBP.

关 键 词：邻苯二甲酸二丁酯 皮层神经元 钙稳态 凋亡 线粒体膜电位 

分 类 号：R114[医药卫生—卫生毒理学]