头孢曲松及其杂质对斑马鱼肝脏的毒性

doi:10.3969/j.issn.1004-616x.2024.01.006

癌变·畸变·突变 ›› 2024, Vol. 36 ›› Issue (1): 35-41,47.doi: 10.3969/j.issn.1004-616x.2024.01.006

头孢曲松及其杂质对斑马鱼肝脏的毒性

张锐¹, 马媛媛¹, 韩莹¹, 崇小萌², 刘馨妍¹, 谢广云³, 梁一帆³, 姚尚辰², 张靖溥¹

1. 中国医学科学院/北京协和医学院医药生物技术研究所, 北京 100050;
2. 中国食品药品检定研究院化学药品检定所抗生素室, 北京 102629;
3. 中国疾病预防控制中心职业卫生与中毒控制所, 北京 100050

收稿日期:2023-09-06 修回日期:2023-11-07 出版日期:2024-02-19 发布日期:2024-02-19
通讯作者: 姚尚辰, 张靖溥
作者简介:张锐,E-mail:zhangrui6166@126.com。
基金资助:
国家“重大新药创制”科技重大专项资助项目（2017ZX09101001-007-003）

Toxicity of ceftriaxone and its impurities to the liver of zebrafish

ZHANG Rui¹, MA Yuanyuan¹, HAN Ying¹, CHONG Xiaomeng², LIU Xinyan¹, XIE Guangyun³, LIANG Yifan³, YAO Shangchen², ZHANG Jingpu¹

1. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050;
2. Division of Antibiotics, Institute for Chemical Drug Control, National Institutes for Food and Drug Control, Beijing 102629;
3. National Institute for Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention, Beijing 100050, China

Received:2023-09-06 Revised:2023-11-07 Online:2024-02-19 Published:2024-02-19

摘要/Abstract

摘要： 目的：研究头孢曲松及其杂质对斑马鱼肝脏的毒性。方法：选用受精后72 h的野生型斑马鱼和肝脏特异性荧光标记的转基因斑马鱼作为实验动物。分别用不同浓度的（0、1、2、5 mmol/L）头孢曲松和杂质A、B、C、D，以及不同浓度的（0、0.1、0.5、1 mmol/L）杂质E处理两种幼鱼2 d后，观察幼鱼肝脏形态和肝脏荧光强度；采用整体油红O染色观察野生型斑马鱼肝脏脂肪含量变化；进一步利用转录组测序技术对各受试物处理组斑马鱼进行转录组测序，筛选差异表达基因，并进行信号通路富集分析。结果：活体观察显示，与对照组比较，头孢曲松及其杂质A和C主要使斑马鱼幼鱼肝脏区扩大或荧光强度增强（P<0.05或0.01），杂质B、D和E主要使幼鱼肝脏区减小或荧光强度减弱（P<0.05或0.01）。整体油红O染色显示头孢曲松及其杂质均能导致斑马鱼肝脏脂肪堆积。头孢曲松给药组利用转录组测序筛选出差异表达基因共735个，杂质A组共237个，杂质C组共237个。KEGG通路分析提示各受试物组差异表达基因主要富集通路不同。头孢曲松差异基因主要富集于代谢通路和卟啉等通路中；杂质A组差异基因主要富集于色氨酸代谢等信号通路；杂质C组差异基因主要富集于钙信号通路等信号通路。结论：头孢曲松及其杂质A、B、C、D和E可导致不同程度斑马鱼肝功能发生变化，并造成肝脏组织损伤。

关键词: 头孢曲松, 肝毒性, 斑马鱼, 差异表达基因, 杂质

Abstract: OBJECTIVE: The objective of this study was to investigate the hepatotoxicity of ceftriaxone and its impurities in zebrafish. METHODS: Wild-type strain zebrafish at 72 h post-fertilization(hpf) and transgenic zebrafish Tg(fabp10a:Ds Red) with liver-specific fluorescence labeling were chosen as experimental animals.Zebrafish larvae were treated with different concentrations of ceftriaxone(at concentrations of 1, 2, 5 mmol/L) and its impurities A, B, C, D and impurities E(at concentrations of 0.1, 0.5, 1 mmol/L) for 2 days. The liver morphology of wild-type zebrafish larvae and the fluorescence intensity of transgenic zebrafish larvae were observed, and the hepatotoxicity of each group was assessed by comparison with the control group. Whole-body Oil Red O staining was employed to observe changes in liver fat content. Additionally, transcriptomic sequencing was performed to detect the gene expression profiles of zebrafish in each treatment group. Differential expression genes were screened(735 differential genes in ceftriaxone group, 237 in impurity A group and 237 in impurity C group), and Gene and Genomes(KEGG) pathway enrichment analysis was conducted. RESULTS: Ceftriaxone and impurities A and C caused an enlargement of the zebrafish liver region or an increase in fluorescence intensity compared to the control group(P<0.05 or 0.01). Impurities B, D, and E primarily resulted in a reduction of the liver region or a decrease in fluorescence intensity compared to the control group(P<0.05 or 0.01). Overall Oil Red O stain assays indicated that both ceftriaxone and its impurities could cause an increase in liver fat. Differential gene expression was observed in each treatment group through transcriptome sequencing.KEGG pathway analysis revealed different pathway enrichments in each group. Genes related to ceftriaxone were mainly enriched in 10 pathways, including metabolism. Genes related to impurity A were mainly enriched in signaling pathways such as tryptophan metabolism. Genes related to impurity C were mainly enriched in signaling pathways, including calcium signaling. CONCLUSION: The study provides insights into the hepatotoxic effects of ceftriaxone and its impurities in zebrafish, demonstrating varied impacts on liver morphology, fluorescence intensity, and gene expression profiles. The findings highlight potential pathways through which these substances may induce hepatotoxicity.

Key words: ceftriaxone, hepatotoxicity, zebrafish, differentially expressed genes, impurity

中图分类号:

R114

张锐, 马媛媛, 韩莹, 崇小萌, 刘馨妍, 谢广云, 梁一帆, 姚尚辰, 张靖溥. 头孢曲松及其杂质对斑马鱼肝脏的毒性[J]. 癌变·畸变·突变, 2024, 36(1): 35-41,47.

ZHANG Rui, MA Yuanyuan, HAN Ying, CHONG Xiaomeng, LIU Xinyan, XIE Guangyun, LIANG Yifan, YAO Shangchen, ZHANG Jingpu. Toxicity of ceftriaxone and its impurities to the liver of zebrafish[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2024, 36(1): 35-41,47.

参考文献

[1] 范琦琦,李芝奇,陈美琳,等.基于斑马鱼模型的中药安全性评价研究进展[J].中草药, 2022, 53(1):278-287.
[2] 孙智慧,贾顺姬,孟安明.斑马鱼:在生命科学中畅游[J].生命科学, 2006, 18(5):431-436.
[3] 陈博,张靖溥.果糖诱导斑马鱼肝脏脂肪变性的转录组学分析[J].中国医药生物技术, 2019, 14(4):308-315.
[4] 陈宁,黄权华,苏军权,等.阿奇霉素及其杂质J对斑马鱼毒性的转录组学分析[J].中国抗生素杂志, 2022, 47(3):241-244.
[5] 刘艺,王瑞昕,游龙泰,等.芦荟大黄素诱导斑马鱼肝毒性及作用机制研究[J].环球中医药, 2020, 13(1):18-22.
[6] 师帅,吴春,陆萍,等.头孢菌素类抗生素的药理特性及临床应用分析[J].中外女性健康研究, 2019(3):32, 68.
[7] 国家药典委员会.中华人民共和国药典-四部:2020年版[M].北京:中国医药科技出版社, 2020.
[8] 耿悦,张锦琳,袁耀佐.国产注射用头孢曲松钠质量分析[J].中南药学, 2020, 18(3):489-492.
[9] 夏青,韩利文,张云,等.基于斑马鱼模型的柴胡皂苷a保肝作用与肝毒性研究[J].中国中药杂志, 2019, 44(13):2662-2666.
[10] 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(3):256-261.
[11] HUANG L, LIU J P, LI W B, et al. Lenvatinib exposure induces hepatotoxicity in zebrafish via inhibiting Wnt signaling[J].Toxicology, 2021, 462:152951.
[12] JIA Z, ZHAO C J, WANG M S, et al. Hepatotoxicity assessment of Rhizoma Paridis in adult zebrafish through proteomes and metabolome[J]. Biomedecine Pharmacother, 2020, 121:109558.
[13] WHO Expert Committee. The selection and use of essential medicines. Report of the WHO expert committee, 2005(including the 14th model list of essential medicines)[J]. World Health Organ Tech Rep Ser, 2006(933):1-119, backcover.
[14] GAO Y, GUO L, HAN Y, et al. A combination of in silico ADMET prediction, in vivo toxicity evaluation, and potential mechanism exploration of brucine and brucine N-oxide-a comparative study[J]. Molecules, 2023, 28(3):1341.
[15] SUN M, LIU Q, LIANG Q X, et al. Toosendanin triggered hepatotoxicity in zebrafish via inflammation, autophagy, and apoptosis pathways[J]. Comp Biochem Physiol C Toxicol Pharmacol, 2021, 250:109171.
[16] 国瑞贤,谢广云,韩莹.基于转录组测序方法研究加替沙星对小鼠的肝损伤作用[J].癌变·畸变·突变, 2022, 34(1):20-24.

头孢曲松及其杂质对斑马鱼肝脏的毒性

Toxicity of ceftriaxone and its impurities to the liver of zebrafish

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	赵立然, 卜梁. 生物信息学分析食管癌关键基因的表达及其临床意义[J]. 癌变·畸变·突变, 2023, 35(5): 374-381.
[2]	谭明雪, 柳晓玲, 姜姝芸, 潘心红, 陈雯, 陈丽萍. 二甲双胍对镉诱导小鼠急性肝毒性的影响[J]. 癌变·畸变·突变, 2023, 35(1): 1-8.
[3]	国瑞贤, 谢广云, 韩莹. 基于转录组测序方法研究加替沙星对小鼠的肝损伤作用[J]. 癌变·畸变·突变, 2022, 34(1): 20-24.
[4]	杨婵君, 陈雯恬, 刘静. 胶原蛋白作为胃癌潜在生物标志物的生物信息学分析[J]. 癌变·畸变·突变, 2021, 33(6): 410-419.
[5]	黄立利, 张梦云, 刘科亮, 庞定国, 刘丽达, 徐培渝. 2,4-二氯苯氧乙酸对幼龄SD大鼠肝毒性的研究[J]. 癌变·畸变·突变, 2021, 33(1): 22-27.
[6]	曾蕾, 徐雪莲, 罗曼曼, 张凡, 韩志坚, 李玉民. 胃癌基因表达谱芯片差异基因的生物信息学分析[J]. 癌变·畸变·突变, 2019, 31(4): 300-307.
[7]	杜太峰, 郑树楷, 黄苑妮, 包冕, 吴库生. 草甘膦对斑马鱼生殖发育的毒性研究进展[J]. 癌变·畸变·突变, 2018, 30(5): 410-412.
[8]	吴德生, 王宏辉, 袁建辉, 王宏菊, 赵琼辉, 刘建军. 细颗粒物对斑马鱼胚胎发育的影响[J]. 癌变·畸变·突变, 2018, 30(4): 307-309,314.
[9]	申碧羚, 郭小玲, 何云. CCM3基因敲降斑马鱼模型的建立及其在毒理学中的初步应用[J]. 癌变·畸变·突变, 2017, 29(3): 194-198,204.
[10]	赖怡, 陈佳红, 张航, 乐聪, 姜岩, 陈涛. 丙烯酰胺对F344大鼠肝毒性的分子机制研究[J]. 癌变·畸变·突变, 2016, 28(3): 174-178.
[11]	李旭玲, 吴德生, 洪文旭, 刘建军, 柯跃斌, 袁建辉, 黎双飞. 纳米SiO₂颗粒对斑马鱼行为变化的影响[J]. 癌变·畸变·突变, 2015, 27(6): 467-471,476.
[12]	李岩，赵英会，庄东明，李晓霞，于爱莲. 脱氧雪腐镰刀菌烯醇致小鼠畸形骨骼组织差异表达基因及相关通路分析[J]. 癌变·畸变·突变, 2015, 27(1): 1-5.
[13]	殷健，王爱平，李万芳，靳洪涛，魏金锋. 一种氧化应激生物标志物定量分析系统的建立及应用[J]. 癌变·畸变·突变, 2014, 26(5): 321-329.
[14]	李建国,秦秀军,袁慧,张伟,周晋源,闻建华. γ射线诱导大鼠外周血淋巴细胞差异表达基因及相关通路的研究[J]. 癌变·畸变·突变, 2013, 25(6): 435-438.
[15]	张伟，秦秀军，许超琪，李炜宾，尹晶晶，袁慧，李建国. γ射线照后不同时间点人肝细胞基因差异表达谱的研究[J]. , 2011, 23(6): 416-420.