哺乳期多溴联苯醚-153染毒对成年大鼠皮质细胞内钙离子浓度和钙激活相关酶的影响  

Effects of polybrominated diphenyl ether-153 lactation exposure on the concentrations of intracellular calcium ion and calcium-activated related enzymes levels of adult rats＇ cerebral cortex

作　　者：张红梅[1] 李新[2] 常利军[1] 张华俊[1] 牛侨[1] 

机构地区：[1]山西医科大学公共卫生学院,太原030001 [2]太原钢铁集团疾病预防控制中心

出　　处：《中华劳动卫生职业病杂志》2013年第7期515-518,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金（81001258）;山西省优秀青年学术带头人（晋教财[20111205号）;山西省青年科技研究基金（2013021033-3）

摘　　要：目的研究哺乳期多溴联苯醚-153（BDE-153）染毒对成年大鼠皮质细胞内钙离子浓度和钙激活相关酶水平的影响，为研究多溴联苯醚的神经发育毒性提供科学依据。方法40只雄性新生乳鼠按体重和窝别随机分成1、5、10mg／kgBDE-153组和橄榄油溶剂对照组，每组10只。在出生第10天，染毒组按0．1ml／10g体重-次腹腔注射染毒BDE-153，橄榄油溶剂对照组给予同剂量橄榄油。染毒后2个月时处死大鼠，冰皿上迅速分离大脑皮质，用流式细胞仪测定细胞内caz＋浓度，用比色法测定钙调神经磷酸酶（CaN）、Ca，Mg2-ATP酶活力，实时荧光定量PCR法和蛋白免疫印迹法（Western．blot）检测钙激活蛋白酶calpain-1和calpain-2的基因和蛋白表达。结果对照组、1、5和10mg／kgBDE-153组大鼠皮质细胞内caz＋平均荧光强度分别为10．83、1．48、1．93和0．62，各染毒组细胞内caz＋浓度均明显低于对照组，差异有统计学意义（P〈O．05）。各染毒组CaN酶活力、Ca2＋-Mg2_ATP酶活力、calpain-1基因和蛋白表达量与对照组的差异无统计学意义（P〉0．05）。随着BDE．153染毒剂量的增加，BDE．15染毒组calpain-2蛋白的表达量逐渐升高，与对照组f基因：（0．81±0．26），蛋白：（0．15±0．07）]比较，5、10mg／kgBDE-153组calpain-2的基因[5mg／l（gBDE-153组：1．16±0．52，10mg，kgBDE．153组：1．32±0．23]及蛋白表达量[5mg／kgBDE．153组：0．31±0．07，10mg／kgBDE-153组：0．37±0．061明显升高，差异有统计学意义（P〈0．05）；10mg／kgBDE．153组calpain-2的蛋白表达量（0．37±0．06）明显高于1mg／kgBDE-153组（0．22±0．07），差异有统计学意义（P（0．05）。结论ca：价导的calpain-2激活可能是BDE-153神经毒性的主要机制之-。Objective To investigate the effects of polybrominated diphenyl ether-153 （BDE-153） exposure during lactation period on the calcium ion （Ca2＋） concentration and calcium-activated enzyme levels in cerebral cortical cells among adult rats and to provide a scientific basis for the study on the developmental neu- rotoxicity of BDE-153. Methods Forty newborn male rats were randomly and equally divided into four groups according to their body weights and litters： 1, 5, and 10 mg/kg BDE-153 groups and olive oil solvent control group. On postnatal day 10 （PND 10）, the BDE-153 groups were administrated BDE-153 （0.1 ml/10 g body weight） by intraperitoneal injection, while the olive oil solvent control group was given an equal volume of olive oil. Two months later, these rats were decapitated, and the cerebral cortex was separated quickly on an ice-cold dish. The Ca2~ concentration in cerebral cortical cells was measured by flow cytometry. The activities of cal- cineurin （CAN） and CaE＋-Mg2＋-ATP enzyme were determined by colorimetric method. The mRNA and protein expression of calpain-1 and calpain-2 was measured by real-time quantitative PCR and Western blot. Results The mean fluorescence intensities of intracellular Ca2＋ in control group and 1, 5, and 10 mg/kg BDE-153 groups were 10.83, 1.48, 1.93, and 0.62, respectively; the 1, 5, and 10 mg/kg BDE-153 groups had significantly lower intercellular Ca2＋ concentrations than the control group （P〈0.05）. The activities of CaN and Ca2＋-Mg＋-ATP enzyme and mRNA and protein expression of calpain-1 showed no significant differences between the 1, 5, and 10 mg/kg BDE-153 groups and control group （P〉0.05）. The protein expression of calpain-2 increased as the dose of BDE- 153 rose. Compared with the control group （mRNA： 0.81＋0.26; protein： 0.15＋0.07）, the 5 and 10 mg/kg BDE-153 groups had significantly higher mRNA expression of calpain-2 （5 mg/kg BDE-153 group： 1.16±0,52; 10 mg/kgBDE-153 group： 1.32±0.23） and significa

关 键 词：多溴联苯醚一153 钙离子 钙激活蛋白酶-1 钙激活蛋白酶 2 钙调神经磷酸酶 caMgATP酶 

分 类 号：R114[医药卫生—卫生毒理学]