| 108 | 0 | 29 |
| 下载次数 | 被引频次 | 阅读次数 |
目的 通过网络毒理学探讨三氧化二砷(ATO)的肝损伤机制，并结合体外实验进行验证。方法 利用DrugBank、CTD、GeneCards、HERB、TTD数据库获取ATO致肝毒性潜在靶点，通过STRING、DAVID数据库结合Cytoscape软件进行蛋白质-蛋白质相互作用(PPI)网络构建、GO富集分析和KEGG通路分析，系统性预测ATO致肝毒性作用机制。通过CCK-8、Fluo-4AM钙离子检测、流式细胞术及Western blot评估AML-12肝细胞增殖、钙稳态、凋亡及内质网应激和JNK/p53通路蛋白表达，并采用SP600125(JNK抑制剂)和PFT-α(p53抑制剂)验证关键通路。结果 筛选出31个核心靶点(如TP53、AKT1)，GO和KEGG分析显示其显著富集于凋亡、癌症等通路。ATO以剂量依赖性抑制细胞增殖，诱导钙超载及内质网应激标志蛋白上调。Western blot证实ATO激活IRE1α-JNK-p53轴，通过调控Bcl-2/Bax平衡诱导线粒体凋亡，JNK/p53抑制剂预处理可显著抑制凋亡。结论 ATO通过“钙稳态失衡-内质网应激-JNK/p53级联”轴介导肝损伤，靶向干预该通路为临床防控提供新策略。
Abstract：Objective To determine the mechanism of arsenic trioxide (ATO)-induced hepatotoxicity via network toxicology and in vitro experiments.Methods Potential targets of ATO-induced hepatotoxicity were identified with the DrugBank,CTD,GeneCards,HERB,and TTD databases.Protein-protein interaction (PPI) network construction,Gene Ontology (GO) enrichment analysis,and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were conducted with STRING,DAVID,and Cytoscape software to predict the mechanism for hepatotoxicity.In vitro experiments assessed the proliferation,calcium homeostasis,apoptosis,endoplasmic reticulum stress markers,and JNK/p53 pathway protein expression in AML-12 hepatocytes with CCK-8 assay,Fluo-4AM calcium detection,flow cytometry,and Western blot.The key pathways were validated by the JNK inhibitor (SP600125) and the p53 inhibitor (PFT-α).Results Thirty-one core targets (e.g.,TP53,AKT1) were identified.GO/KEGG analysis revealed obvious enrichment in apoptosis and cancer-related pathways.ATO dose-dependently inhibited the cell proliferation,induced calcium overload,and upregulated endoplasmic reticulum stress markers.Western blot demonstrated that ATO selectively activated the IRE1α-JNK-p53 axis,thereby driving the mitochondrial apoptosis via dysregulation of the Bcl-2/Bax ratio.Pretreatment with JNK/p53 inhibitors much attenuated the apoptosis.Conclusion ATO mediates hepatocyte injury through the calcium dyshomeostasis-endoplasmic reticulum stress-JNK/p53 cascade axis,providing a novel therapeutic target for clinically preventing and managing ATO-induced hepatotoxicity.
[1]Abdul KSM，Jayasinghe SS，Chandana EPS，et al.Arsenic and human health effects：a review[J].Environ Toxicol Pharmacol，2015，40(3)：828-846.
[2]Paul NP，Galván AE，Yoshinaga-Sakurai K，et al.Arsenic in medicine：past，present and future[J].Biometals，2023，36(2)：283-301.
[3]张亭栋，张鹏飞，王守仁，等.“癌灵注射液”治疗6例白血病初步临床观察[J].黑龙江医药，1973(3)：66-67.
[4]孙鸿德，马玲，胡晓晨，等.癌灵 Ⅰ 号结合中医辨证治疗急性早幼粒细胞白血病32例[J].中国中西医结合杂志，1992，12(3)：170-171.
[5]中华医学会血液学分会，中国医师协会血液科医师分会.中国急性早幼粒细胞白血病诊疗指南(2018年版)[J].中华血液学杂志，2018，39(3)：179-183.
[6]Levis M.Arsenic and all-trans retinoic acid for acute promyelocytic leukemia：yes，it really is as good as it seems[J].Haematologica，2021，106(12)：3031-3032.
[7]张玲玲，丰志培，王良悦，等.基于传承创新的中医药与现代科学结合路径探析——以砷剂治疗白血病为例[J].中南药学，2023，21(10)：2704-2709.
[8]Zhang Z，Zhang SJ，Zhang F，et al.Clinical indicators of hepatotoxicity in newly diagnosed acute promyelocytic leukemia patients undergoing arsenic trioxide treatment[J].Biol Trace Elem Res，2024，202(1)：122-132.
[9]Gunes EK，Ozkurt ZN，Yildiz S，et al.Comparison of maintenance regimens in acute promyelocytic leukemia patient[J].Leuk Res，2023，128：107055.
[10]Liu GZ，Song YR，Li CX，et al.Arsenic compounds：the wide application and mechanisms applied in acute promyelocytic leukemia and carcinogenic toxicology[J].Eur J Med Chem，2021，221：113519.
[11]Huang QC，Chen YW，Zhang ZJ，et al.The endoplasmic reticulum participated in drug metabolic toxicity[J].Cell Biol Toxicol，2022，38(6)：945-961.
[12]Loh SN.Arsenic and an old place：rescuing p53 mutants in cancer[J].Cancer Cell，2021，39(2)：140-142.
[13]Singh AP，Goel RK，Kaur T.Mechanisms pertaining to arsenic toxicity[J].Toxicol Int，2011，18(2)：87-93.
[14]刘春燕，张爱华.内质网应激与p53在NaAsO2诱导肝细胞凋亡中的作用[C]//中国毒理学会.中国毒理学会第十次全国毒理学大会论文集.贵州医科大学环境污染与疾病监控教育部重点实验室公共卫生与健康学院，2023：52-53.
[15]Renu K，Saravanan A，Elangovan A，et al.An appraisal on molecular and biochemical signalling cascades during arsenic-induced hepatotoxicity[J].Life Sci，2020，260：118438.
[16]Liu YQ，Stockwell BR，Jiang XJ，et al.p53-regulated non-apoptotic cell death pathways and their relevance in cancer and other diseases[J].Nat Rev Mol Cell Biol，2025，26(8)：600-614.
[17]Fusée L，Salomao N，Ponnuswamy A，et al.The p53 endoplasmic reticulum stress-response pathway evolved in humans but not in mice via PERK-regulated p53 mRNA structures[J].Cell Death Differ，2023，30：1072-1081.
[18]Vogler M，Braun Y，Smith VM，et al.The BCL2 family：from apoptosis mechanisms to new advances in targeted therapy[J].Signal Transduct Target Ther，2025，10(1)：91.
[19]Zhang XY，Yang SM，Zhang HP，et al.Endoplasmic reticulum stress mediates the arsenic trioxide-induced apoptosis in human hepatocellular carcinoma cells[J].Int J Biochem Cell Biol，2015，68：158-165.
[20]Wang RY，Zhang JY，Wang S，et al.The cardiotoxicity induced by arsenic trioxide is alleviated by salvianolic acid A via maintaining calcium homeostasis and inhibiting endoplasmic reticulum stress[J].Molecules，2019，24(3)：543.
[21]蓝天伟，于倩.肉苁蓉苯乙醇苷拮抗H2O2诱发H9c2细胞内质网应激损伤的保护作用[J].中南药学，2018，16(10)：1383-1387.
[22]Zhao L，Wang LY，Wang H，et al.Endoplasmic reticulum chaperone GRP78 participates in fluoride-induced autophagy in cells by regulating the IRE1-TRAF2-JNK pathway[J].Environ Toxicol，2023，38(7)：1756-1767.
[23]牟艳卉，陈昭桂，姚光，等.针刺通过活性氧/c-Jun氨基末端激酶/p53通路对创伤后应激障碍大鼠模型海马神经元凋亡的影响机制[J].实用临床医药杂志，2025，29(1)：44-49.
[24]Rozpedek W，Pytel D，Mucha B，et al.The role of the PERK/eIF2α/ATF4/CHOP signaling pathway in tumor progression during endoplasmic reticulum stress[J].Curr Mol Med，2016，16(6)：533-544.
[25]Akhter N，Wilson A，Arefanian H，et al.Endoplasmic reticulum stress promotes the expression of TNF-α in THP-1 cells by mechanisms involving ROS/CHOP/HIF-1α and MAPK/NF-κB pathways[J].Int J Mol Sci，2023，24(20)：15186.
基本信息:
中图分类号:R99
引用信息:
[1]黄庆财,巨珊珊,白秀云,等.三氧化二砷通过钙稳态失衡-内质网应激-JNK/p53级联引起AML-12肝细胞损伤机制分析[J].中南药学,2026,24(03):69-75.
基金信息:
国家重点研发计划(No.19YFC1709200)
2025-05-25
2025
2026-03-26
2026
2026-01-09
1
2026-03-19
2026-03-19