双酚S经ERβ-MAPK信号通路介导对人正常卵巢上皮细胞的毒性作用  

Toxicity of bisphenol S on human normal ovarian epithelial cells mediated by ERβ-MAPK signaling pathway

作　　者：李晓蒙 于梦淇 田林 熊围 张立实 杨芷蕊 陈锦瑶 Li Xiaomeng;Yu Mengqi;Tian Lin;Xiong Wei;Zhang Lishi;Yang Zhirui;Chen Jinyao(Department of Nutrition and Food Hygiene,West China School of Public Health/West China Fourth Hospital/Food Safety Monitoring and Risk Assessment Key Laboratory of Sichuan Province,Sichuan University,Chengdu 610041,China;Business Management and Information Technology Department,Chengdu Institute of Food Inspection,Chengdu 611130,China)

机构地区：[1]四川大学华西公共卫生学院/华西第四医院营养与食品卫生学系/食品安全监测与风险评估四川省重点实验室,成都610041 [2]成都市食品检验研究院业务管理与信息技术部,成都611130

出　　处：《卫生研究》2024年第3期441-446,454,共7页Journal of Hygiene Research

基　　金：国家重点研发计划(No.2017YFC1600201)。

摘　　要：目的探讨模拟人体实际暴露水平的双酚S长期(7和14 d)染毒,通过ERβ-MAPK信号通路扰动,对人正常卵巢上皮细胞IOSE 80的毒性效应。方法使用基于生理的药代动力学模型结合中国长江沿岸地区女性血清双酚S水平确定给药浓度,同时设置溶剂对照和β-雌二醇(17β-estradiol,E_(2))对照,对IOSE 80细胞进行长期染毒(7和14 d),每2天更换一次相应浓度(0、6.79×10^(-6)、6.79×10^(-4)、6.79×10^(-2)、6.79μmol/L双酚S和10 nmol/L E_(2))完全培养基。通过qPCR检测雌激素受体(ERβ和GPR30)、MAPK信号通路(P38/JNK/ERK信号通路)关键基因和促性腺激素释放激素相关基因(GnRH-I、GnRH-II和GnRH-R)mRNA表达;在此基础上给予ERβ-MAPK信号通路抑制剂,采用流式细胞术检测细胞周期;同时进行剂量-反应关系分析,计算相关指标的基准剂量可信限下限值。结果与溶剂对照组相比,双酚S染毒7 d后,G_(2)/M期细胞比例显著增加(P<0.05),GnRH-I、GnRH-II和GnRH-R的mRNA表达量显著下调(P<0.05);双酚S染毒14 d后,ESR2、MAPK3、MAPK9的mRNA表达量显著上调(P<0.05),GnRH-II和GnRH-R的mRNA表达量显著下调(P<0.05)。双酚S染毒7 d的GnRH-II mRNA表达量,双酚S染毒14 d的G_(0)/G_(1)期比例、MAPK3和MAPK8 mRNA表达量,以及双酚S染毒7和14 d后的GnRH-I mRNA表达量,呈现较好的剂量-反应关系,但拟合结果较差。结论双酚S长期低剂量染毒可激活ERβ-MAPK信号通路引起IOSE 80细胞周期阻滞,也可导致IOSE 80细胞激素分泌变化。OBJECTIVE To investigate the effects of long-term(7 days and 14 days)bisphenol S(BPS)exposure on the ERβ-MAPK signaling pathway,hormone secretion phenotype and cell cycle in human normal ovarian epithelial cells IOSE 80 at actual human exposure level.METHODS Physiologically based pharmacokinetic model combined with BPS levels in the serum of women along the Yangtze River in China was used to determine the dosing concentrations of BPS,and vehicle control and 17β-estradiol(E_2)control were used.Complete medium with corresponding concentrations(0,6.79×10^(-6),6.79×10^(-4),6.79×10^(-2),6.79μmol/L BPS and 10 nmol/L E_(2))was replaced every 2 days.mRNA expressions of estrogen receptor(ERβand GPR30),key genes in MAPK signaling pathway(P38/JNK/ERK signaling pathway)and gonadotropin-releasing hormone-related genes(GnRH-I,GnRH-II and GnRH-R)were measured by qPCR.The ERβ-MAPK signaling pathway inhibitors were employed to detect the effect of long-term exposure to BPS on the cell cycle by flow cytometry.Dose-response relationship analysis was performed to calculate the benchmark does lower confidence limits.RESULTSCompared to the vehicle control,after 7 days exposure to BPS,the ratio of G_(2)/M phase was significantly increased(P<0.05),and the mRNA expressions of GnRH-I,GnRH-II and GnRH-R were significantly decreased(P<0.05);after 14 days exposure to BPS,the mRNA expressions of ESR2,MAPK3,and MAPK9 were significantly increased(P<0.05),and the mRNA expressions of GnRH-II and GnRH-R were significantly decreased(P<0.05).The GnRH-II mRNA expression level of BPS treatment for 7 days;the G_(0)/G_(1)phase ratio,MAPK3 and MAPK8 mRNA expression level of BPS exposure for 14 days;and the GnRH-I mRNA expression level after BPS treatment for 7 days and 14 days showed a good dose-response relationship but with poor fit.CONCLUSION Long-term low-dose exposure to BPS may cause cell cycle arrest by activating the ERβ-MAPK signaling pathway,and may lead to changes in the hormone secretion of IOSE 80 cells.

关 键 词：双酚S 人正常卵巢上皮细胞 雌激素受体 MAPK信号通路 基准剂量 

分 类 号：R155.5[医药卫生—营养与食品卫生学]