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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

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徐旻昊, 镇华君, 修光利. 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[J]. 生态毒理学报, 2024, 19(6): 12-20. doi: 10.7524/AJE.1673-5897.20240607001
引用本文: 徐旻昊, 镇华君, 修光利. 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[J]. 生态毒理学报, 2024, 19(6): 12-20. doi: 10.7524/AJE.1673-5897.20240607001
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XU Minhao, ZHEN Huajun, XIU Guangli. Investigation on Effects of Typical OPFRs on CREB Transcriptional Activity and Adipogenesis During Early Differentiation of Mouse Preadipocyte 3T3-L1[J]. Asian journal of ecotoxicology, 2024, 19(6): 12-20. doi: 10.7524/AJE.1673-5897.20240607001
Citation: XU Minhao, ZHEN Huajun, XIU Guangli. Investigation on Effects of Typical OPFRs on CREB Transcriptional Activity and Adipogenesis During Early Differentiation of Mouse Preadipocyte 3T3-L1[J]. Asian journal of ecotoxicology, 2024, 19(6): 12-20. doi: 10.7524/AJE.1673-5897.20240607001
shu

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

  • 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

    • 作者简介: 徐旻昊(1999—)，男，硕士，研究方向为环境毒理学，E-mail:sss709212547@163.com
    通讯作者: 镇华君(1985-),男,博士,副教授,主要研究方向为污染物的毒理和健康效应。E-mail:zhenhuajun@ecust.edu.cn;  修光利(1972—)，男，博士，教授，主要研究方向为大气环境化学、大气污染控制以及环境风险评价与规划管理。E-mail:xiugl@ecust.edu.cn
  • 基金项目:

    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)

  • 中图分类号: X171.5

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

  • 1. School of Resources and Environmental Engineering, East China University of Science and Technology, Shanghai 200237, China;

  • 2. Shanghai Environmental Protection Key Laboratory on Environmental Standard and Risk Management of Chemical Pollutants, Shanghai 200237, China;

  • 3. State Environmental Protection Key Laboratory of Environmental Risk Assessment and Control on Chemical Process, Shanghai 200237, China;

  • 4. Shanghai Institute of Pollution Control and Ecological Security, Shanghai 200092, China

    • Corresponding authors: ZHEN Huajun ;  XIU Guangli
  • Fund Project:

  • 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10 μmol·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10 μmol·L-1的6种OPFRs(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。
    Abstract: 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 10 μmol·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.
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  • Yao C, Yang H P, Li Y. A review on organophosphate flame retardants in the environment: Occurrence, accumulation, metabolism and toxicity[J]. Science of the Total Environment, 2021, 795: 148837
    Ali N, Ali L, Mehdi T, et al. Levels and profiles of organochlorines and flame retardants in car and house dust from Kuwait and Pakistan: Implication for human exposure via dust ingestion[J]. Environment International, 2013, 55: 62-70
    Hu W X, Gao P, Wang L, et al. Endocrine disrupting toxicity of aryl organophosphate esters and mode of action[J]. Critical Reviews in Environmental Science and Technology, 2023, 53(1): 1-18
    Dishaw L V, Powers C M, Ryde I T, et al. Is the Penta-BDE replacement, tris (1,3-dichloro-2-propyl) phosphate (TDCPP), a developmental neurotoxicant? Studies in PC12 cells[J]. Toxicology and Applied Pharmacology, 2011, 256(3): 281-289
    Wang Q W, Lam J C W, Man Y C, et al. Bioconcentration, metabolism and neurotoxicity of the organophorous flame retardant 1,3-dichloro 2-propyl phosphate (TDCPP) to zebrafish[J]. Aquatic Toxicology, 2015, 158: 108-115
    Li H, Li F L, Zhou C Y, et al. Exposure to OPFRs is associated with obesity and dysregulated serum lipid profiles: Data from 2017-2018 NHANES[J]. Metabolites, 2024, 14(2): 124
    Yang D Q, Wei X J, Zhang Z Y, et al. Tris (2-chloroethyl) phosphate (TCEP) induces obesity and hepatic steatosis via FXR-mediated lipid accumulation in mice: Long-term exposure as a potential risk for metabolic diseases[J]. Chemico-Biological Interactions, 2022, 363: 110027
    Wang D Z, Yan S, Yan J, et al. Effects of triphenyl phosphate exposure during fetal development on obesity and metabolic dysfunctions in adult mice: Impaired lipid metabolism and intestinal dysbiosis[J]. Environmental Pollution, 2019, 246: 630-638
    Liu Q S, Sun Z D, Ren X M, et al. Chemical structure-related adipogenic effects of tetrabromobisphenol A and its analogues on 3T3-L1 preadipocytes[J]. Environmental Science & Technology, 2020, 54(10): 6262-6271
    Li C H, Ren X M, Guo L H. Adipogenic activity of oligomeric hexafluoropropylene oxide (perfluorooctanoic acid alternative) through peroxisome proliferator-activated receptor γ pathway[J]. Environmental Science & Technology, 2019, 53(6): 3287-3295
    Zebisch K, Voigt V, Wabitsch M, et al. Protocol for effective differentiation of 3T3-L1 cells to adipocytes[J]. Analytical Biochemistry, 2012, 425(1): 88-90
    Poulos S P, Dodson M V, Hausman G J. Cell line models for differentiation: Preadipocytes and adipocytes[J]. Experimental Biology and Medicine, 2010, 235(10): 1185-1193
    Pillai H K, Fang M L, Beglov D, et al. Ligand binding and activation of PPARγ by Firemaster® 550: Effects on adipogenesis and osteogenesis in vitro[J]. Environmental Health Perspectives, 2014, 122(11): 1225-1232
    Tung E W Y, Ahmed S, Peshdary V, et al. Firemaster® 550 and its components isopropylated triphenyl phosphate and triphenyl phosphate enhance adipogenesis and transcriptional activity of peroxisome proliferator activated receptor (PPARγ) on the adipocyte protein 2(aP2) promoter[J]. PLoS One, 2017, 12(4): e0175855
    Yue J J, Sun C T, Tang J Y, et al. Downregulation of miRNA-155-5p contributes to the adipogenic activity of 2-ethylhexyl diphenyl phosphate in 3T3-L1 preadipocytes[J]. Toxicology, 2023, 487: 153452
    Kassotis C D, Hoffman K, Stapleton H M. Characterization of adipogenic activity of house dust extracts and semi-volatile indoor contaminants in 3T3-L1 cells[J]. Environmental Science & Technology, 2017, 51(15): 8735-8745
    Wang X Y, Sun Z D, Liu Q S, et al. Environmental obesogens and their perturbations in lipid metabolism[J]. Environment & Health, 2024, 2(5): 253-268
    Farmer S R. Transcriptional control of adipocyte formation[J]. Cell Metabolism, 2006, 4(4): 263-273
    Reusch J E, Colton L A, Klemm D J. CREB activation induces adipogenesis in 3T3-L1 cells[J]. Molecular and Cellular Biology, 2000, 20(3): 1008-1020
    Zhang J W, Klemm D J, Vinson C, et al. Role of CREB in transcriptional regulation of CCAAT/enhancer-binding protein beta gene during adipogenesis[J]. Journal of Biological Chemistry, 2004, 279(6): 4471-4478
    Mayr B M, Canettieri G, Montminy M R. Distinct effects of cAMP and mitogenic signals on CREB-binding protein recruitment impart specificity to target gene activation via[WT《Times New Roman#I》] CREB[J]. Proceedings of the National Academy of Sciences of the United States of America, 2001, 98(19): 10936-10941
    Sun Z D, Yang X X, Liu Q S, et al. Butylated hydroxyanisole isomers induce distinct adipogenesis in 3T3-L1 cells[J]. Journal of Hazardous Materials, 2019, 379: 120794
    Delghandi M P, Johannessen M, Moens U. The cAMP signalling pathway activates CREB through PKA, p38 and MSK1 in NIH 3T3 cells[J]. Cellular Signalling, 2005, 17(11): 1343-1351
    Hua Y J, Ke S S, Wang Y, et al. Prolonged treatment with 3-isobutyl-1-methylxanthine improves the efficiency of differentiating 3T3-L1 cells into adipocytes[J]. Analytical Biochemistry, 2016, 507: 18-20
    Nunes A R, Batuca J R, Monteiro E C. Acute hypoxia modifies cAMP levels induced by inhibitors of phosphodiesterase-4 in rat carotid bodies, carotid arteries and superior cervical Ganglia[J]. British Journal of Pharmacology, 2010, 159(2): 353-361
    Laurenza A, Sutkowski E M, Seamon K B. Forskolin: A specific stimulator of adenylyl cyclase or a diterpene with multiple sites of action?[J]. Trends in Pharmacological Sciences, 1989, 10(11): 442-447
    Gonzalez-Iglesias A E, Jiang Y H, Tomić M, et al. Dependence of electrical activity and calcium influx-controlled prolactin release on adenylyl cyclase signaling pathway in pituitary lactotrophs[J]. Molecular Endocrinology, 2006, 20(9): 2231-2246
    Sun W J, Duan X Y, Chen H, et al. Adipogenic activity of 2-ethylhexyl diphenyl phosphate via peroxisome proliferator-activated receptor γ pathway[J]. Science of the Total Environment, 2020, 711: 134810
    Zhang Q, Wang J H, Zhu J Q, et al. Potential glucocorticoid and mineralocorticoid effects of nine organophosphate flame retardants[J]. Environmental Science & Technology, 2017, 51(10): 5803-5810
    Kojima H, Takeuchi S, Itoh T, et al. In vitro endocrine disruption potential of organophosphate flame retardants via human nuclear receptors[J]. Toxicology, 2013, 314(1): 76-83
    Skledar D G, Carino A, Trontelj J, et al. Endocrine activities and adipogenic effects of bisphenol AF and its main metabolite[J]. Chemosphere, 2019, 215: 870-880
    Hao Z L, Zhang Z J, Lu D Z, et al. Organophosphorus flame retardants impair intracellular lipid metabolic function in human hepatocellular cells[J]. Chemical Research in Toxicology, 2019, 32(6): 1250-1258
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  • 收稿日期:  2024-06-07
徐旻昊, 镇华君, 修光利. 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[J]. 生态毒理学报, 2024, 19(6): 12-20. doi: 10.7524/AJE.1673-5897.20240607001
引用本文: 徐旻昊, 镇华君, 修光利. 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[J]. 生态毒理学报, 2024, 19(6): 12-20. doi: 10.7524/AJE.1673-5897.20240607001
shu
XU Minhao, ZHEN Huajun, XIU Guangli. Investigation on Effects of Typical OPFRs on CREB Transcriptional Activity and Adipogenesis During Early Differentiation of Mouse Preadipocyte 3T3-L1[J]. Asian journal of ecotoxicology, 2024, 19(6): 12-20. doi: 10.7524/AJE.1673-5897.20240607001
Citation: XU Minhao, ZHEN Huajun, XIU Guangli. Investigation on Effects of Typical OPFRs on CREB Transcriptional Activity and Adipogenesis During Early Differentiation of Mouse Preadipocyte 3T3-L1[J]. Asian journal of ecotoxicology, 2024, 19(6): 12-20. doi: 10.7524/AJE.1673-5897.20240607001
shu

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

    通讯作者: 镇华君(1985-),男,博士,副教授,主要研究方向为污染物的毒理和健康效应。E-mail:zhenhuajun@ecust.edu.cn;  修光利(1972—)，男，博士，教授，主要研究方向为大气环境化学、大气污染控制以及环境风险评价与规划管理。E-mail:xiugl@ecust.edu.cn
    作者简介: 徐旻昊(1999—)，男，硕士，研究方向为环境毒理学，E-mail:sss709212547@163.com
  • 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
  • 收稿日期:  2024-06-07
基金项目:

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10 μmol·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10 μmol·L-1的6种OPFRs(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。

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