典型有机磷阻燃剂对小鼠前脂肪细胞3T3-L1的CREB转录活性及成脂分化影响研究  

Investigation on Effects of Typical OPFRs on CREB Transcriptional Activity and Adipogenesis During Early Differentiation of Mouse Preadipocyte 3T3-L1

作　　者：徐旻昊 镇华君 修光利[1,2,3,4] XU Minhao;ZHEN Huajun;XIU Guangli(School of Resources and Environmental Engineering,East China University of Science and Technology,Shanghai 200237,China;Shanghai Environmental Protection Key Laboratory on Environmental Standard and Risk Management of Chemical Pollutants,Shanghai 200237,China;State Environmental Protection Key Laboratory of Environmental Risk Assessment and Control on Chemical Process,Shanghai 200237,China;Shanghai Institute of Pollution Control and Ecological Security,Shanghai 200092,China)

机构地区：[1]华东理工大学资源与环境工程学院,上海200237 [2]上海市环境保护化学污染物环境标准与风险管理重点实验室,上海200237 [3]国家环境保护化工过程环境风险评价与控制重点实验室,上海200237 [4]上海污染控制与生态安全研究院,上海200092

出　　处：《生态毒理学报》2024年第6期12-20,共9页Asian Journal of Ecotoxicology

基　　金：国家自然科学基金面上项目(22276056);上海市浦江人才计划资助项目(20PJ1402700)。

摘　　要：有机磷阻燃剂(organophosphate flame retardants,OPFRs)具有诱导小鼠前脂肪细胞3T3-L1分化为成熟脂肪细胞的能力,并促使细胞积累大量的甘油三酯(triglyceride,TG),因此被视为潜在的致肥胖污染物。目前,关于OPFRs诱导3T3-L1成脂分化的作用机制研究尚不充分,特别是在细胞分化早期阶段(诱导后0~2 d)污染物的影响作用仍然不明确。环磷腺苷效应元件结合蛋白(cAMP response element-binding protein,CREB)是3T3-L1分化早期的关键调控因子。为了探究OPFRs在诱导0~2 d暴露对CREB转录活性的影响,本研究构建了基于环磷腺苷效应元件(cAMP response element,CRE)和pGL4.73质粒共转染的双荧光素酶报告基因方法。进一步对10种典型的OPFRs开展检测,发现10μmol·L^(-1)的2-乙基己基二苯基磷酸酯(EHDPP)、磷酸二(2-乙基己基)苯基酯(BEHPP)、磷酸三(1,3-二氯-2-丙基)酯(TDCIPP)、磷酸三(2-氯乙基)酯(TCEP)、磷酸三(2-乙基己基)酯(TEHP)可显著上调分化早期3T3-L1细胞的CREB转录活性。此外,在诱导分化的0~2 d内,暴露于10μmol·L^(-1)的6种O PFRs(BEHPP、EHDPP、TEHP、TDCIPP、磷酸三(2-氯异丙基)酯(TCIPP)、TCEP)可显著增加细胞分化结束后的TG含量,且TG的生成量与分化早期CREB转录活性存在边缘显著水平(P=0.07)的正相关关系。本研究揭示了不同取代基结构的OPFRs对3T3-L1成脂分化早期阶段的差异化调控作用,增进了对OPFRs致肥胖效应的认识,并为评估OPFRs健康风险提供了重要的科学依据。Organophosphate flame retardants(OPFRs)possess the capability to induce the differentiation of mouse preadipocytes 3T3-L1 into mature adipocytes,leading to a significant accumulation of triglycerides(TG)within the cells.Consequently,they might be considered as potential obesogens.Currently,the mechanism by which OPFRs induce adipogenic differentiation in 3T3-L1 cells is not well understood,particularly regarding the impact of pollutants during the early stage of cell differentiation(0-2 days post-induction).Cyclic AMP response element-binding protein(CREB)is a key regulator in the early stages of 3T3-L1 differentiation.To investigate the impact of OPFRs exposure on CREB transcriptional activity,this study developed a dual-luciferase reporter gene assay based on co-transfection with cAMP response element(CRE)and pGL4.73 plasmid.Further testing of 10 typical OPFRs revealed that,2-ethylhexyl diphenyl phosphate(EHDPP),bis(2-ethylhexyl)phenyl phosphate(BEHPP),tris(1,3-dichloro-2-propyl)phosphate(TDCIPP),tris(2-chloroethyl)phosphate(TCEP),and tris(2-ethylhexyl)phosphate(TEHP)significantly upregulated CREB transcriptional activity in 3T3-L1 cells at a concentration of 10μmol·L^(-1) during the early stages of differentiation,respectively.Moreover,the TG contents in cells exposed to six OPFRs(BEHPP,EHDPP,TEHP,TDCIPP,tris(2-chloroisopropyl)phosphate(TCIPP),TCEP)for first two days were found to significantly increase when differentiation was completed.Notably,there was a marginally significant positive correlation(P=0.07)between TG production and CREB transcriptional activity in the early stages of differentiation.The present study reveals the differential regulatory effects of OPFRs with different substituent structures in the early stages of 3T3-L1 differentiation,improves current understanding of the obesogenic effects of OPFRs,and provides an important scientific basis for assessing the health risks of OPFRs.

关 键 词：有机磷阻燃剂 小鼠 致肥胖效应 CREB通路 3T3-L1细胞 

分 类 号：X171.5[环境科学与工程—环境科学]