双酚A抑制大鼠胚胎中脑微团细胞的存活和分化与Notch-Hes通路有关  

Bisphenol A inhibits cell viability and differentiation in micromass culture of rat embryonic midbrain cells via the Notch-Hes pathway

作　　者：刘然[1,2] 蒋建军[1] 尚兰琴[1] 魏雪涛[1] 吴双[1] 郝卫东[1] 

机构地区：[1]北京大学医学部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 [2]农业部农药检定所,北京100125

出　　处：《中国药理学与毒理学杂志》2014年第2期175-181,共7页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金(30771821);国家自然科学基金(30972501)~~

摘　　要：目的探讨雌激素受体(ER)途径在双酚A(BPA)诱导的大鼠胚胎中脑微团细胞存活和分化抑制效应中的作用。方法制备的胎龄13 d大鼠胚胎中脑微团细胞,以BPA 10-4,10-6,10-8,10-10和10-12mol·L-1染毒5 d,中性红摄取实验检测细胞存活,苏木精染色检测中脑细胞集落分化总面积。BPA0.1 mmol·L-1+雌激素受体抑制剂ICI182780 0.1 nmol·L-1、BPA 0.1 mmol·L-1+雌激素受体抑制剂他莫西芬1 nmol·L-1共同培养5 d,检测中脑微团细胞存活和集落分化总面积。Western blot法检测胎龄18 d大鼠胚胎脑组织、成年大鼠的睾丸和未染毒中脑微团细胞ER蛋白的表达。实时定量PCR法检测胎龄13和18 d胎鼠脑组织、成年大鼠的睾丸及卵巢、未染毒中脑微团细胞中ER mRNA的表达水平。采用实时定量PCR法检测BPA 0.1 mmol·L-1染毒5 d的中脑微团细胞中Notch1和Hes1 mRNA表达水平。结果 BPA 0.1 mmol·L-1可以明显的抑制原代培养中脑微团细胞的存活和集落分化,但此效应不能被ICI182780和他莫西芬逆转。正常的胎鼠脑组织和未染BPA中脑细胞中ER蛋白和mRNA的表达水平较低。BPA0.1 mmol·L-1可以明显增加Notch1和Hes1 mRNA表达水平。结论 BPA可抑制中脑微团细胞的的存活和分化,ER途径在此效应中不发挥主要作用,Notch-Hes信号通路可能参与了这一效应。OBJECTIVE Study the role of estrogen receptor （ER）in the inhibition of cell viability and differentiation induced by bisphenol A （BPA）in micro mass culture of rat e mbryonic midbrain（MB） cells.METHODS Micro mass cultures of MB were prepared fro m rat e mbryonic midbrain on gestation day 13.MB cells were exposed to BPA （10 -4 ,10 -6 ,10 -8 ,10 -10 ,10 -12 mol·L -1 ）for 5 d.Cell viability was assessed by neutral red uptake test.MB differentiation was detected by he matoxylin staining and i mage analysis.In order to observe the role of ER pathway in the toxicity induced by BPA,cell cultures were co-treated with ICI182780 0.1 n mol·L -1 ,ta moxifen 1 n mol·L -1 and BPA 0.1 mmol·L -1 for 5 d, the cell viability and foci differentiation were detected.Moreover,the protein expression levels of ER in normal e mbryonic brain of gestation day 18,testis tissue fro m adult rats and midbrain cells untreated with BPA were investigated by Western blot.The mRNA expression levels of ER in normal e mbryonic brain of gestation day 13 and gestation day 18,ovary and testis tissue fro m adult rats,and midbrain cells un-treated with BPA were investigated by real-ti me PCR.The mRNA expression levels of Notch1 and Hes1 in MB cells treated with BPA 0.1 mmol·L -1 were also detected by real-ti me PCR.RESULTS BPA 0.1 mmol·L -1 could inhibited MB cell viability and foci differentiation.However,this effect could not be reversed by ER antagonist.The protein and mRNA expression levels of ER in e mbryonic brain and MB cells untreated with BPA were found to be extre mly low.In addition,BPA 0.1 mmol·L -1 could inhibited the mRNA expression levels of Notch1 and Hes1 .CONCLUSION BPA could inhibited MB cell viability and foci differentiation.ER pathway might be not involved in this effect.Instead,Notch-Hes pathway might be involved for this effect.

关 键 词：双酚A 微团培养 雌激素受体 细胞存活 细胞分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]