恩诺沙星在鲫鱼肝微粒体中的代谢及代谢关键酶
Metabolism of Enrofloxacin in Liver Microsomes of Crucian Carp (Carassius auratus) and Its Key Enzymes in vitro
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摘要: 抗生素因具有抗菌谱广、杀菌性强等特点而被广泛应用于人类医药、畜牧业、农业和水产养殖业。其进入水生生物体内后，会在药物代谢酶的作用下发生代谢转化，产生生态毒性。采用鲫鱼肝微粒体体外孵育法，探究恩诺沙星细胞色素P450酶作用下的代谢转化过程，并通过代谢抑制实验确定关键的代谢酶。结果表明，恩诺沙星体外代谢过程符合一级动力学方程，当恩诺沙星暴露浓度为1 mg·L-1时，其在肝微粒体中的消除速率常数（k）最大为0.00303 min-1，半衰期（t1/2）最短为228.8 min，应用高效液相色谱串联质谱（HPLC-MS/MS）技术，检测到了恩诺沙星脱乙基产物和羟基化产物；代谢抑制实验结果表明，CYP3A4在恩诺沙星代谢过程中起主要作用，是恩诺沙星代谢的关键酶。本研究结果为深入了解恩诺沙星在水生生物体内的代谢转化及其生态风险提供了基础数据。Abstract: Enrofloxacin is efficient and broad-spectrum, which makes it widely used in human medicine, animal husbandry, agriculture and aquaculture. Enrofloxacin can be metabolized by drug metabolizing enzymes in aquatic organisms, which will affect its ecological toxicity. In this study, the in vitro incubation method of liver microsomes was used to explore the metabolic transformation process of enrofloxacin under the action cytochrome P450 enzyme, and the key metabolic enzymes were determined by metabolic inhibition experiments. The results showed that the metabolic process of enrofloxacin in fish liver microsomes met the first-order kinetic equation, the maximum depuration rate constant (k) of enrofloxacin in the fish liver microsomes was 0.00303 min-1 and the minimum half-life (t1/2) was 228.8 min in 1 mg·L-1 of enrofloxacin exposure concentration. High performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was used to identify the metabolites and metabolic pathways of enrofloxacin in liver microsomes. Two major metabolites were found, including the product of deethylation and hydroxylation. Further, this metabolic inhibition study showed that cytochrome P3A4 might be the key enzyme involved in the metabolism of enrofloxacin. Overall, this study provides basic data for further understanding of the biotransformation and ecological risk of enrofloxacin in aquatic organisms.
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Key words:
- enrofloxacin /
- Carassius auratus /
- microsomes /
- in vitro metabolism /
- CYP3A4
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牛曰华, 张天闻, 邹红梅, 等. 恩诺沙星及其代谢物环丙沙星在大黄鱼体内的代谢动力学[J]. 中国渔业质量与标准, 2018, 8(1):24-33 Niu Y H, Zhang T W, Zou H M, et al. Pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin in Larimichthys crocea[J]. Chinese Fishery Quality and Standards, 2018, 8(1):24-33(in Chinese)
陈红英, 王月颖, 傅思武. 抗生素在养殖业中的应用现状[J]. 现代畜牧科技, 2019(5):1-3 Chen H Y, Wang Y Y, Fu S W. Application status of antibiotics in aquaculture[J]. Modern Animal Husbandry Science & Technology, 2019 (5):1-3(in Chinese)
王立红, 刘芳, 管振国, 等. 偏碱性恩诺沙星注射液对大鼠亚慢性毒性试验研究[J]. 天津科技, 2017, 44(7):49-52 Wang L H, Liu F, Guan Z G, et al. Subchronic toxicity of alkali deviation type enrofloxacin injection in SD rats[J]. Tianjin Science & Technology, 2017, 44(7):49-52(in Chinese)
Guengerich F P. Cytochrome P450 and chemical toxicology[J]. Chemical Research in Toxicology, 2008, 21(1):70-83 Fan J, Shan Q, Wang J, et al. Comparative pharmacokinetics of enrofloxacin in healthy and aeromonas hydrophila-infected crucian carp (Carassius auratus gibelio)[J]. Journal of Veterinary Pharmacology and Therapeutics, 2017, 40(5):580-582 Morales-Gutierrez F J, Barbosa J, Barron D. Metabolic study of enrofloxacin and metabolic profile modifications in broiler chicken tissues after drug administration[J]. Food Chemistry, 2015, 172:30-39 Ruennarong N, Wingpanit K, Sakulthaew C, et al. Dispositions of enronfloxacin and its major metabolite ciprofloxacin in Thai swamp buffaloes[J]. Journal of Veterinary Medical Science, 2016, 78(3):397-403 Bonassa K P D, Miragliotta M Y, Simas R C, et al. Tissue depletion study of enrofloxacin and its metabolite ciprofloxacin in broiler chickens after oral administration of a new veterinary pharmaceutical formulation containing enrofloxacin[J]. Food & Chemical Toxicology, 2017, 105:8-13 沈梦楠. 典型溴代阻燃剂在鲫鱼体内与体外代谢研究[D]. 南京:南京大学, 2012:51 Shen M N. Metabolism of typical brominated flame retardants by crucian carp in vivo and in vitro[D]. Nanjing:Nanjing University, 2012:51(in Chinese) 周帅, 李国烈, 胡琳琳, 等. 恩诺沙星在异育银鲫体内和体外肝微粒体中代谢产物分析[J]. 东北农业大学学报, 2013, 44(6):101-106 Zhou S, Li G L, Hu L L, et al.In vivo and liver microsome in vitro analysis of metabolites of enrofloxacin from crucian carp (Carassius auratus gibelio) using UPLC/Q-TOF MS[J]. Journal of Northeast Agricultural University, 2013, 44(6):101-106(in Chinese)
Burkina V, Zlabek V, Rasmussen M K, et al. End-product inhibition of skatole-metabolising enzymes CYP1A, CYP2A19 and CYP2E1 in porcine and piscine hepatic microsomes[J]. Toxicology Letters, 2019, 303:67-71 李阳, 史雪岩, 高希武. 增效醚对二化螟幼虫7-乙氧基香豆素-O-脱乙基酶和羧酸酯酶活性的影响[J].昆虫学报, 2016, 59(11):1159-1165 Li Y, Shi X Y, Gao X W. In vivo effects of piperonyl butoxide on the activities of 7-ethoxycoumarin O-deethylase and carboxylesterase in larval Chilo suppressalis (Lepidoptera:Pyralidae)[J]. Acta Entomologica Sinica, 2016, 59(11):1159-1165(in Chinese)
Shan Q, Fan J. Pharmacokinetics of enrofloxacin after oral, intramuscular and crucian carp (Carassius auratus gibelio)[J]. Journal of Veterinary Pharmacology and Therapeutics, 2018, 41(1):159-162 Hansen J, Palmfeldt J, Pedersen K W, et al. Postmortem protein stability investigations of the human hepatic drug-metabolizing cytochrome P450 enzymes CYP1A2 and CYP3A4 using mass spectrometry[J]. Journal of Proteomics, 2019, 194:125-131 Uwimana E, Ruiz P, Li X S, et al. Human CYP2A6, CYP2B6, and CYP2E1 atropselectively metabolize polychlorinated biphenyls to hydroxylated metabolites[J]. Environmental Science & Technology, 2019, 53(4):2114-2123 陈曦, 王建书, 姜英, 等. B(a)P和DEHP联合染毒对HepG2细胞CYP1A1和CYP3A4酶活性的影响[J].环境与职业医学, 2011, 28(9):525-530 Chen X, Wang J S, Jiang Y, et al. Effects of co-exposure of di-(2-ethylhexyl) phthalate and benzo (a) pyrene on CYP1A1 and CYP3A4 enzyme activities in HepG2 cells[J]. Journal of Environmental & Occupational Medicine, 2011, 28(9):525-530(in Chinese)
Hou R, Huang C, Rao K F, et al. Characterized in vitro metabolism kinetics of alkyl organophosphate esters in fish liver and intestinal microsomes[J]. Environmental Science & Technology, 2018, 52(5):3202-3210 Shen M N, Cheng J, Wu R H, et al. Metabolism of polybrominated diphenyl ethers and tetrabromobisphenol A by fish liver subcellular fractions in vitro[J]. Aquatic Toxicology, 2012, 114-115:73-79 李文, 麦曦, 胡晓. 1-(4-溴苄基)-1-(4-溴苄氧基)脲药代动力学和体外代谢的研究[J]. 中国临床药理学杂志, 2013, 29(9):675-678 Li W, Mai X, Hu X. Pharmacokinetics and metabolism of 1-(4-bromobenzyl)-1-(4-bromobenzyloxy) urea in vitro[J]. The Chinese Journal of Clinical Pharmacology, 2013, 29(9):675-678(in Chinese)
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