张琪^1，， 吴双^1，*， 刘璐^2，， 侯晓鸿^1，， 蒋建军^1，， 魏雪涛^1，， 郝卫东^1，

（  1、北京大学公共卫生学院毒理系，食品安全毒理学研究与评价北京市重点实验室，北京 100191；       2、北京大学基础医学院医学遗传学系，北京 100191；  ）

摘要： 目的：研究双酚A（BPA）对人类正常皮肤细胞HaCat细胞间缝隙连接通讯（GJIC）的影响和可能机制。方法：采用改良E-Screen法评估细胞增殖情况，采用荧光漂白后恢复技术（FRAP）评估GJIC功能，采用实时荧光定量PCR检测mRNA转录水平。结果：BPA在1.0×10-8-1.0×10-6 mol/L可显著提高HaCat细胞增殖率，延长荧光恢复时间，降低荧光恢复率。雌激素受体抑制剂ICI182780可部分阻断BPA的上述作用，ERK信号通路抑制剂U0126可完全抑制BPA的促增殖作用，却不能挽救BPA对GJIC的抑制。BPA 1.0×10-7mol/可显著下调Cx26 mRNA转录水平，ICI182780和U0126均可抑制此下调作用。BPA1.0×10-7，1.0×10-6 mol/L在与U0126共同作用时，能够显著上调ERK1/2 mRNA转录水平；BPA1.0×10-7mol/L与ICI182780共同作用时，结果类似U0126但较弱。结论：BPA能通过ER-ERK信号通路刺激HaCat 细胞增殖，能通过雌激素受体（ER）途径抑制HaCat细胞GJIC功能，能通过ER-ERK信号通路下调Cx26 mRNA转录水平。ERK通路的完全抑制可激发BPA促进ERK1/2 mRNA转录，且ER在其中发挥重要作用。
关键词： 卫生毒理学；双酚A；HaCat；缝隙连接；雌激素受体途径；ERK信号通路

ZHANG Qi^1,， WU Shuang^1,*， LIU Lu^2,， HOU Xiaohong^1,， JIANG Jianjun^1,， WEI Xuetao^1,， HAO Weidong^1,

（  1、Department of Toxicology, School of Public Health, Peking University, Beijing Key Laboratory of Toxicological Research and Risk Assessment for Food Safety, Beijing 100191；       2、Department of Genetics, School of Basic Medical Science, Peking University, Beijing 100191；  ）

Abstract： Objective: The aim of this study was to investigate the effects and possible mechanisms of BPA on gap junctional intercellular communication (GJIC) of human normal skin cells HaCat. Methods: A modified E-Screen assay was used to evaluate cell proliferation, fluorescence recovery after photobleaching (FRAP) technique was used to evaluate GJIC function and Real-time Fluorescent Quantitative PCR was used to detect mRNA expression levels. Results: BPA increased cell proliferation rates, prolonged fluorescence recovery times, and reduced fluorescence recovery rates at 1.0×10-8-1.0×10-6 mol/L. Estrogen receptor (ER) inhibitor ICI182780 partially blocked the above effects. MAPK/ERK kinase (MEK) 1/2 inhibitor U0126 completely blocked the promoting effects on cell proliferation, but did not rescue the inhibition of GJIC. BPA 1.0×10-7 mol/L downregulated Cx26 mRNA level. ICI182780 and U0126 inhibited this effect. BPA 1.0×10-7 and 1.0×10-6 mol/L combined with U0126 up-regulated extracellular regulated protein kinases (ERK) 1/2 mRNA levels. BPA 1.0×10-7 mol/L combined with ICI182780 had similar but weaker effects. Conclusion: BPA can promote HaCat proliferation through ER-ERK pathway, inhibit HaCat GJIC function through ER pathway, and downregulate Cx26 mRNA level through ER-ERK pathway. Complete inhibition of ERK pathway can stimulate BPA to promote ERK 1/2 mRNA expression, and ER plays an important role in this effect.
Keywords： Healthtoxicology; Bisphenol A; HaCat; Gap junction; Estrogen receptor pathway; ERK signaling

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