2,7-二溴咔唑致肝损伤分子机制的网络毒理学辅助实验探讨  

Network toxicology-assisted experiments to explore 2,7-dibromocarbazole's hepatotoxic mechanism

作　　者：张巧云 姚敦璠 汪国威 侯存闯 付强 黄梦瑶 ZHANG Qiaoyun;YAO Dunfan;WANG Guowei;HOU Cunchuang;FU Qiang;HUANG Mengyao(Key Laboratory for Green Chemical Process of Ministry of Education,School of Environmental Ecology and BiologicalEngineering,Wuhan Institute of Technology,Wuhan 430205,China;School of Pharmacy,Xianning Medical College,Hubei University of Science and Technology,Xianning 437100,China)

机构地区：[1]武汉工程大学绿色化工过程教育部重点实验室,环境生态与生物工程学院,湖北武汉430205 [2]湖北科技学院医学部药学院,湖北咸宁437100

出　　处：《武汉工程大学学报》2024年第5期534-542,共9页Journal of Wuhan Institute of Technology

基　　金：国家自然科学基金(21806047、41902314);湖北省教育厅科学基金(Q20211512);武汉工程大学研究生教育创新基金(CX2022567)。

摘　　要：环境污染物2,7-二溴咔唑(27-BCZ)显著引发肝脏毒性效应,研究其分子机制将在生态风险评估和毒性靶向治疗药物的开发中起到重要作用。证实0.05和0.15μmol/L的27-BCZ可致斑马鱼肝损伤,引起肝脏ALT和AST水平升高,肝脏部分细胞溶解、胞核固缩和变性等。网络毒理学方法预测显示:27-BCZ致肝损伤的关键靶点为RXRα、MAPK1、HSP90AA1等,其机制主要涉及PI3K-Akt信号通路。分子对接表明27-BCZ与PI3K-Akt信号通路相关的核心靶点RXRα结合活性强。为验证网络毒理学预测的可靠性,用RT-qPCR检测肝组织PI3K-Akt信号通路介导凋亡相关基因的表达水平,结果显示:27-BCZ可显著降低肝脏pi3k、akt和bcl-2基因的表达水平,升高凋亡相关的bax和caspase 3基因的表达水平,这种降低或升高的趋势随着暴露浓度的升高进一步增强。综合预测与实验结果发现,27-BCZ致肝损伤可能与其下调肝脏PI3K-Akt信号通路引发肝细胞凋亡有关。本研究首次采用网络毒理学方法探究环境污染物的潜在肝脏毒性作用机制,为环境有机污染物毒性机制的研究提供了新的思路与方法。The environmental pollutant 2,7-dibromocarbazole(27-BCZ)significantly induces liver toxicity,thus identification of its molecular mechanism on liver toxicity play a vital role in ecological risk assessment and the development of therapeutics.This study confirmed that 0.05μmol/L or 0.15μmol/L 27-BCZ can induce liver damage in zebrafish due to partial cell lysis,nuclear pyknosis and denaturation,and an increase in ALT and AST in the liver.The network toxicology predicted that the key targets of 27-BCZ in inducing liver damage were RXRα,MAPK1,HSP90AA1,etc.,and the mechanisms involved PI3K-Akt signaling pathways.The molecular docking results indicated that the core target RXRαassociated with the PI3K-Akt signaling pathway had strong binding activity with 27-BCZ.To verify the feasibility of network toxicology prediction,RT-qPCR was used to detect apoptosis-related genes mediated by the PI3K-Akt signaling pathway in the liver.The results showed that 27-BCZ significantly reduced the transcriptional levels of genes pi3k,akt,and bcl-2,and significantly increased transcriptional levels of genes bax and caspase 3 which are related to cell apoptosis.The trends of decrease or increase were further enhanced with increasing concentration of 27-BCZ.Based on the network toxicological and experimental results,we found that 27-BCZ-induced liver damage may be related to its downregulation of the PI3K-Akt signaling pathway and promotion of liver cell apoptosis.This is the first study that attempted using the network toxicology methods to explore the hepatic toxicity mechanisms of environmental pollutants,which provides new insight into investigation of the toxicity mechanism of environmental organic pollutants.

关 键 词：卤代咔唑 肝脏毒性 RXRα靶点 PI3K-AKT信号通路 

分 类 号：X131[环境科学与工程—环境科学] R99[医药卫生—毒理学]