Abstract：In this study, zebrafish served as a model organism to meticulously investigate the toxicological effects of azithromycin (AZM) on larval development during acute exposure. Our research delineated several distinct pathological changes in the larvae, including spinal curvature, incomplete swim bladder development, and the formation of yolk cysts. Notably, profound neurological impacts were also observed, characterized by a significant reduction in the number of neurons within the zebrafish brain, diminished AChE activity, and disruptions in light-dark rhythms. Further investigation using the PharmMapper website identified potential AZM target genes. Subsequent GO and KEGG enrichment analysis unveiled the potential influence of AZM on zebrafish vascular and immune systems. Experimental validation substantiated these predictions, demonstrating that AZM exposure induces abnormal vascular branching and dysfunction in the immune system of zebrafish larvae.
[1] Aydin S, Aydin M E, Ulvi A, et al. Antibiotics in hospital effluents:occurrence, contribution to urban wastewater, removal in a wastewater treatment plant, and environmental risk assessment[J]. Environmental Science and Pollution Research, 2019,26(1):544-558. [2] Danner M C, Robertson A, Behrends V, et al. Antibiotic pollution in surface fresh waters:Occurrence and effects[J]. Science of the Total Environment, 2019,664:793-804. [3] Ostman M, Lindberg R H, Fick J, et al. Screening of biocides, metals and antibiotics in Swedish sewage sludge and wastewater[J]. Water Research, 2017,115:318-328. [4] Wang J, Wei H, Zhou X, et al. Occurrence and risk assessment of antibiotics in the Xi'an section of the Weihe River, northwestern China[J]. Marine Pollution Bulletin, 2019,146:794-800. [5] Duong Q A, Pittet L F, Curtis N, et al. Antibiotic exposure and adverse long-term health outcomes in children:A systematic review and meta-analysis[J]. Journal of Infection, 2022,85(3):213-300. [6] Rodrigues S, Antunes S C, Nunes B, et al. Histological alterations in gills and liver of rainbow trout (Oncorhynchus mykiss) after exposure to the antibiotic oxytetracycline[J]. Environmental Toxicology and Pharmacology, 2017,53:164-176. [7] Shiogiri N S, Ikefuti C V, Carraschi S P, et al. Effects of azithromycin on tilapia (Oreochromis niloticus):health status evaluation using biochemical, physiological and morphological biomarkers[J]. Aquaculture Research, 2017,48(7):3669-3683. [8] Qiu W H, Liu T, Liu X J, et al. Enrofloxacin Induces Intestinal Microbiota-Mediated Immunosuppression in Zebrafish[J]. Environmental Science & Technology, 2022,56(12):8428-8437. [9] Qiu W H, Chen B, Tang L, et al. Antibiotic Chlortetracycline Causes Transgenerational Immunosuppression via NF-kappa B[J]. Environmental Science & Technology, 2022,56(7):4251-4261. [10] Li R, Wang H X, Shi Q F, et al. Effects of oral florfenicol and azithromycin on gut microbiota and adipogenesis in mice[J]. Plos one, 2017,12(7):e0181690. [11] 张苗青,张靖溥.红霉素和阿奇霉素诱导斑马鱼肝毒性及作用机制研究[J]. 中国抗生素杂志, 2021,46:256-261. Zhang M Q, Zhang J P. Study on hepatotoxicity inducement and mechanisms of erythromycin and azithromycin in zebrafish[J]. Chinese Journal of Antibiotics, 2021,46:256-261. [12] 崔国祯,周鹤峰,陈焕娴,等.阿奇霉素对斑马鱼心脏毒性的初步研究[J]. 遵义医学院学报, 2014,37:495-498. Cui G Z, Zhou H F, Chen H X, et al. Pilot study on cardiotoxicity induced by azithromycin in zebrafish embryos[J]. Journal of Zunyi Medical University. 2014,37:495-498. [13] 郭子一,王伟伟,宋杰,等.基于Illumina RNA-Seq分析的三种内分泌干扰物对斑马鱼神经毒性效应与机制[J]. 中国环境科学, 2023,43(2):946-956. Guo Z Y, Wang W W, Song J, et al. Neurotoxicity effects and mechanisms of three endocrine disruptors on zebrafish based on Illumina RNA-seq analysis[J]. China Environmental Science, 2023, 43(2):946-956. [14] 赵晨曦,王杨,钱秋慧,等.三氯卡班环境暴露对斑马鱼神经行为的影响[J]. 中国环境科学, 2022,42(1):456-464. Zhao C X, Wang Y, Qian Q H, et al. Effects of environmental exposure to triclocarban on the neurobehavior of zebrafish (Danio rerio)[J]. China Environmental Science, 2022,42(1):456-464. [15] Gu J, Guo M, Yin X, et al. A systematic comparison of neurotoxicity of bisphenol A and its derivatives in zebrafish[J]. Science of the Total Environment, 2022,805:150210. [16] Wang D, Wang X, Huang H, et al. Triclosan regulates alternative splicing events of nerve-related genes through RNA-binding protein CELF2 to induce zebrafish neurotoxicity[J]. Journal of Hazardous Materials, 2021,413:125414. [17] Li V W T, Tsui M P M, Chen X, et al. Effects of 4-methylbenzylidene camphor (4-MBC) on neuronal and muscular development in zebrafish (Danio rerio) embryos[J]. Environmental Science and Pollution Research, 2016,23(9):8275-8285. [18] Pullaguri N, Nema S, Bhargava Y, et al. Triclosan alters adult zebrafish behavior and targets acetylcholinesterase activity and expression[J]. Environmental Toxicology and Pharmacology, 2020,75:103311. [19] Cao X, Fu M, Du Q, et al. Developmental toxicity of black phosphorus quantum dots in zebrafish (Danio rerio) embryos[J]. Chemosphere, 2023, 335:139029. [20] Bagwell J, Norman J, Ellis K, et al. Notochord vacuoles absorb compressive bone growth during zebrafish spine formation[J]. eLife, 2020,9:e51221. [21] Coffin A B, Dale E, Doppenberg E, et al. Putative COVID-19therapies imatinib, lopinavir, ritonavir, and ivermectin cause hair cell damage:A targeted screen in the zebrafish lateral line[J]. Frontiers in Cellular Neuroscience, 2022, 24:16:941031. [22] Mendonca-gomes J M, Da costa araujo A P, Da luz T M, et al. Environmental impacts of COVID-19treatment:Toxicological evaluation of azithromycin and hydroxychloroquine in adult zebrafish[J]. Science of the Total Environment, 2021,790:148129. [23] Che X, Huang Y, Zhong K, et al. Thiophanate-methyl induces notochord toxicity by activating the PI3K-mTOR pathway in zebrafish (Danio rerio) embryos[J]. Environmental Pollution, 2023,318:120861. [24] Wang W, Li X, Qian Q, et al. Mechanistic exploration on neurodevelopmental toxicity induced by upregulation of alkbh5targeted by triclosan exposure to larval zebrafish[J]. Journal of Hazardous Materials, 2023,457:131831. [25] Li Q, Lin J, Zhang Y, et al. Differential behavioral responses of zebrafish larvae to yohimbine treatment[J]. Psychopharmacology, 2015,232(1):197-208. [26] Oliveira R, Domingues I, Koppe grisolia C, et al. Effects of triclosan on zebrafish early-life stages and adults[J]. Environmental Science and Pollution Research, 2009,16(6):679-688. [27] Sheng J, Gong J, Shi Y, et al. MicroRNA-22coordinates vascular and motor neuronal pathfinding via sema4during zebrafish development[J]. Open Biology, 2022,12(4):210315. [28] Chen C, Song J, Pu Q, et al. Azithromycin induces neurotoxicity in zebrafish by interfering with the VEGF/Notch signaling pathway[J]. Science of the Total Environment, 2023,903:166505. [29] Tian L, Ma l, Kaarela T, et al. Neuroimmune crosstalk in the central nervous system and its significance for neurological diseases[J]. Journal of Neuroinflammation, 2012,9:155. [30] Var S R, Byrd-Jacobs C A. Role of macrophages and microglia in zebrafish regeneration[J]. International journal of Molecular Sciences, 2020,21(13):4768. [31] Langenau D M, Zon L I. The zebrafish:A new model of T-cell and thymic development[J]. Nature Reviews Immunology, 2005,5(4):307-317.
