基于斑马鱼模型的交泰丸安全性评价

张明哲; 陈章美; 莫柳英; 郝二伟; 侯小涛; 温晓燕; 邓家刚; 杜正彩

doi:10.13422/j.cnki.syfjx.20201821

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基于斑马鱼模型的交泰丸安全性评价

药理 | 更新时间：2020-09-09

- 基于斑马鱼模型的交泰丸安全性评价
- Safety Evaluation of Jiaotaiwan Based on Zebrafish Model
- 中国实验方剂学杂志 2020年26卷第18期页码：51-57
- 作者机构：
  
  1.广西中医药大学科学实验中心，药学院，南宁 530200
  2.多伦多大学医学院，多伦多 M5S 1A8
- 作者简介：
  
  张明哲，在读硕士，从事中药药理与中药新药研发研究，E-mail：zhangmingzhe184@163.com
  杜正彩，硕士，研究员，从事中药药效筛选及新产品开发研究，E-mail：duzhengcai8@163.com
- 基金信息：
  
  广西中医药大学校级重点硕士研究生科研创新项目(YCSZ20190001;YCSZ20190022);广西中药药效研究重点实验室(省级)开放课题项目(17-259-20-P5)
- DOI：10.13422/j.cnki.syfjx.20201821
  中图分类号： R22;R242;R2-031;R285.5
- 收稿日期：2020-07-11，
  
  网络出版日期：2020-07-21，
  
  纸质出版日期：2020-09-20
- 稿件说明：
移动端阅览
张明哲,陈章美,莫柳英等.基于斑马鱼模型的交泰丸安全性评价[J].中国实验方剂学杂志,2020,26(18):51-57.

ZHANG Ming-zhe,CHEN Zhang-mei,MO Liu-ying,et al.Safety Evaluation of Jiaotaiwan Based on Zebrafish Model[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(18):51-57.
张明哲,陈章美,莫柳英等.基于斑马鱼模型的交泰丸安全性评价[J].中国实验方剂学杂志,2020,26(18):51-57. DOI： 10.13422/j.cnki.syfjx.20201821.

ZHANG Ming-zhe,CHEN Zhang-mei,MO Liu-ying,et al.Safety Evaluation of Jiaotaiwan Based on Zebrafish Model[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(18):51-57. DOI： 10.13422/j.cnki.syfjx.20201821.

摘要

目的

以斑马鱼胚胎为研究对象，观察交泰丸水提物对斑马鱼胚胎的生长发育毒性，心脏毒性及肝肾毒性，为交泰丸临床安全合理用药及新产品开发提供参考。

方法

生长发育及心脏毒性实验以受精后12 h（hpf）发育正常的斑马鱼胚胎作为模型动物，以125，250，500 mg·L

-1

交泰丸水提物培养上述胚胎，于受精后72 h（hpf）观察半数致死量及药物对胚胎心率及心脏形态学的影响；肝肾毒性实验以受精后72 h（hpf）发育正常的斑马鱼幼鱼作为模型动物，以125，250，500 mg·L

-1

交泰丸水提物培养上述胚胎，于受精后7 d（dpf）观察半数致死量及药物对斑马鱼幼鱼形态学改变，检测丙氨酸氨基转移酶（ALT），天冬氨酸氨基转移酶（AST）活性和肌酐含量。

结果

生长发育毒性实验中空白组斑马鱼胚胎发育正常，心脏发育完好，心跳均匀有力。交泰丸水提物对斑马鱼胚胎72 h的半数致死量（LD

）为1 023 mg·L

-1

。与空白组胚胎比较，250，500 mg·L

-1

交泰丸水提物组可以观察到胚胎头部变小，体长变短（

0.05），眼睛半径减小（

0.05），可见显著心包水肿，心率显著降低（

0.01），静脉窦-动脉球（SV-BA）间距明显增大（

0.05），心房、心室面积均明显减小（

0.05），瓣膜间距（AV channel）距离显著变大（

0.01），入血口距离显著变小（

0.01）。急性毒性实验中，交泰丸水提物对斑马鱼幼鱼72 h的LD

为1 067 mg·L

-1

；与空白组比较，交泰丸水提物可明显降低斑马鱼幼鱼体内ALT活性（

0.05）。

结论

该实验发现交泰丸具有明显胚胎发育毒性，主要表现为生长发育延缓及严重的心脏毒性，对孕妇、哺乳期妇女及心脏病患者临床用药时应予以高度重视。

Abstract

Objective

To take zebrafish embryo as the research object

in order to investigate the development toxicity

cardiotoxicity

liver toxicity and kidney toxicity of water extract of Jiaotaiwan (JTW) on zebrafish embryo.

Method

Zebrafish embryos with normal development at 12 h (hpf) after fertilization were selected as model animals for the growth and cardiotoxicity experiments. The embryos were treated with 125

250

500 mg·L

-1

of JTW water extracts

and the effects of the drugs on the heart rate and morphology of the embryos and LD

were observed at 72 h (hpf) after fertilization. Zebrafish embryos with normal development at 72 h (hpf) after fertilization were used as model animals for the liver and kidney toxicity experiments. The embryos were treated with 125，250，500 mg·L

-1

of JTW water extracts

and the effect of the drugs on morphological changes

Alanine aminotransferase(ALT)

Aspartate aminotransferase (AST) activity

and creatinine content of the larvae and LD

were observed at 72 d (dpf) after fertilization.

Result

The zebrafish embryos in control group developed normally

the heart was well developed

and the heartbeat was even and powerful. The LD

of JTW water extract on zebrafish embryos for 72 h was 1 023 mg·L

-1

. Compared with the embryos in the control group

250，500 mg·L

-1

treatment groups in the development toxicity had a smaller head

shorter body lengths (

0.05)

and decreased eye size (

0.05). Compared with the control group embryos

the pericardial edema was observed in the 500 mg·L

-1

group

the heart rate was significantly decreased in the 250，500 mg·L

-1

JTW water extract groups (

0.01)

the atrial and ventricular areas were significantly reduced (

0.05)

the distance of SV-BA became significantly larger (

0.05)

the distance of AV channel became significantly larger (

0.01)

and the in-flow distance was significantly shorter (

0.01). In the acute toxicity experiment

the LD

of JTW water extract for zebrafish larvae for 72 h was 1 067 mg·L

-1

. Compared with control group

JTW water extract significantly reduced ALT activity in zebrafish larvae (

0.05).

Conclusion

This experiment found that JTW has an obvious toxicity in embryonic development

which is mainly manifested as delayed growth and severe cardiotoxicity. Great attention shall be paid to clinical administration to pregnant women

lactating women and patients with heart disease.

关键词

Keywords

references

祝建材 . 从交泰丸谈“心肾相交理论” [J]. 中国中医基础医学杂志 , 2017 , 23 ( 1 ): 117 - 118 .

翁剑平 . 交泰丸治疗心肾不交型不寐症的临床疗效研究 [D]. 杭州：浙江中医药大学 , 2019 .

陈志亮 . 斑马鱼在药物筛选中的应用 [J]. 中国中药杂志 , 2015 , 40 ( 7 ): 1235 - 1239 .

李娟娥 , 姜小帆 . 交泰丸对糖尿病小鼠认知功能障碍的影响及机制 [J]. 中国实验方剂学杂志 , 2019 , 25 ( 17 ): 23 - 27 .

王雪萍 , 张皓月 , 王婷 , 等 . 交泰丸中小檗碱联合cinnamtannin D 1 的降血糖作用 [J]. 中成药 , 2018 , 40 ( 12 ): 2613 - 2618 .

雒明池 , 梁如 , 高杉 , 等 . 基于NO-cGMP信号转导通路交泰丸对慢性温和不可预知性应激抑郁大鼠的抗抑郁作用研究 [J]. 中草药 , 2018 , 49 ( 18 ): 4344 - 4348 .

雒明池 , 梁如 , 高树明 , 等 . 基于cAMP-CREB-BDNF信号通路探讨交泰丸抗抑郁的作用机制 [J]. 天津中医药 , 2018 , 35 ( 5 ): 365 - 369 .

GERLAI R , LEE V , BLASER R . Effects of acute and chronic ethanol exposure on the behavior of adult zebrafish (Danio rerio) [J]. Pharmacol Biochem Behav , 2006 , 85 ( 4 ): 752 - 761 .

ZON L I , PETERSON R T . In vivo drug discovery in the zebrafish [J]. Nat Rev Drug Discov , 2005 , 4 ( 1 ): 35 - 44 .

罗隽 , 梅雪 , 夏青 , 等 . 醋甘遂不同提取物对斑马鱼幼鱼的毒性 [J]. 中国实验方剂学杂志 , 2018 , 24 ( 4 ): 160 - 166 .

彭夕洋 , 陈婷芳 , 黄婷 , 等 . 心脏特异表达绿色荧光斑马鱼模型的建立与评估 [J]. 遗传 , 2013 , 35 ( 4 ): 411 - 418 .

LI S , JIANG Y , SUN Q , et al . Tebuconazole induced oxidative stress related hepatotoxicity in adult and larval zebrafish (Danio rerio) [J]. Chemosphere , 2020 , 241 : 125 - 129 .

DING Y J , CHEN Y H . Developmental nephrotoxicity of aristolochic acid in a zebrafish model [J]. Toxicol Appl Pharmacol , 2012 , 261 ( 1 ): 59 - 65 .

DINH Y J , SUN C Y , WEN C C , et al . Nephroprotective role of resveratrol and ursolic acid in aristolochic acid intoxicated zebrafish [J]. Toxins , 2015 , 7 ( 1 ): 97 - 109 .

全云云 , 周忆梦 , 刘美辰 , 等 . 斑马鱼模型筛选何首乌肝毒性的物质基础 [J]. 中国实验方剂学杂志 , 2019 , 25 ( 6 ): 52 - 57 .

全正扬 , 孙震晓 . 基于斑马鱼模型的何首乌水提物及其主要成分的肝毒性研究 [J]. 中国药物警戒 , 2018 , 15 ( 1 ): 1 - 5 .

HILL A J , TERAOKA H , HEIDEMAN W , et al . Zebrafish as a model vertebrate for investigating chemical toxicity [J]. Toxicol Sci , 2005 , 86 ( 1 ): 6 - 19 .

SEGNER H . Zebrafish (Danio rerio) as a model organism for investigating endocrine disruption [J]. Comp Biochem Physiol C Toxicol Pharmacol , 2009 , 149 ( 2 ): 187 - 195 .

曹雨诞 , 张楷承 , 姚芳 , 等 . 京大戟醋制前后对斑马鱼胚胎心脏的毒性 [J]. 中国实验方剂学杂志 , 2019 , 25 ( 24 ): 73 - 77 .

WARREN K S , FISHMAN M C . " Physiological genomics": mutant screens in zebrafish [J]. Am J Physiol , 1998 , 275 ( 1 Pt 2): H1-H7 .

PARNG C , SENG W L , SEMINO C , et al . Zebrafish: a preclinical model for drug screening [J]. Assay Drug Dev Technol , 2002 , 1 ( 1 Pt 1 ): 41 - 48 .

LOMBÓ M , GONZÁLEZ-ROJO S , FERNÁNDEZ-DÍEZ C , et al . Cardiogenesis impairment promoted by bisphenol A exposure is successfully counteracted by epigallocatechin gallate [J]. Environ Pollut ， 2019 , 246 : 1008 - 1019 .

YANG Y X , LI B B , ZHANG X , et al . The zinc finger protein Zfpm1 modulates ventricular trabeculation through Neuregulin-ErbB signalling [J]. Dev Biol ， 2019 , 446 ( 2 ): 142 - 150 .

HUNG M W , ZHANG Z J , LI S , et al . From omics to drug metabolism and high content screen of natural product in zebrafish: a new model for discovery of neuroactive compound [J]. Evid-Based Complement Alternat Med , 2012 , 2012 : 605303 .

张勇 , 贺剑辉 , 刘洪翠 , 等 . 斑马鱼在药物临床前毒理学研究中的应用进展及现状 [J]. 中国药理学与毒理学杂志 , 2013 , 27 ( 3 ): 482 - 483 .

钱拉拉 , 张红 , 翁金月 , 等 . 交泰丸结合针刺治疗心肾不交型失眠的疗效及对血清TNF- α 、IL-6、IL-1 β 的影响 [J]. 中华中医药学刊 , 2019 , 37 ( 3 ): 525 - 527 .

ZHANG M Y , YU Y Y , WANG S F , et al . Cardiotoxicity evaluation of nine alkaloids from, Rhizoma Coptis [J]. Hum Exp Toxicol , 2018 , 37 ( 2 ): 185 - 195 .

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