中国药物警戒 ›› 2019, Vol. 16 ›› Issue (7): 385-392.

• 基础与临床研究 •    下一篇

基于2D和3D肝细胞模型的何首乌体外肝毒性评价

颜玉静1, 淡墨1, 汪祺2,*, 文海若1,*   

  1. 1 中国食品药品检定研究院国家药物安全评价监测中心,药物非临床安全评价研究北京市重点实验室,北京 100176;
    2 中国食品药品检定研究院中药民族药检定所,北京 100050
  • 收稿日期:2019-08-06 修回日期:2019-08-06 出版日期:2019-07-20 发布日期:2019-08-06
  • 通讯作者: 汪祺,女,博士,副研究员,中药毒理。E-mail：sansan8251@sina.com;文海若,女,博士,副研究员,药物安全性评价。E-mail：wenhairuo@nifdc.org.cn
  • 作者简介:颜玉静,女,在读硕士,药物安全性评价。
  • 基金资助:
    国家自然科学基金资助项目（81503347,81503068）：基于体内外代谢产物及UGT1A1抑制作用探索何首乌肝毒性机制及毒效物质基础,氧化应激通过TRPC受体介导心律失常作用机制的研究; 国家十三五“重大新药创制”专项（2018ZX09201017）：创新药物非临床安全性评价关键技术研究

Hepatotoxicity Evaluation of Polygonum Multiflorum Based on 2D and 3D Hepatocyte Models In Vitro

YAN Yujing1, DAN Mo1, WANG Qi2,*, WEN Hairuo1,*   

  1. 1 National Center for Safety Evaluation of Drugs, National Institutes for Food and Drug Control, Key Laboratory of Beijing for Nonclinical Safety Evaluation Research of Drugs, Beijing 100176, China;
    2 Institute for Control of Chinese Traditional Medicine and Ethnic Medicine, National Institutes for Food and Drug Control, Beijing 100050, China
  • Received:2019-08-06 Revised:2019-08-06 Online:2019-07-20 Published:2019-08-06

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摘要： 目的 分别使用二维（two-dimension,2D）和三维（three-dimension,3D）培养的人源肝细胞模型评价何首乌醇提物（polygonum multiflorum ethanol extract,PMEE）和2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷（2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside,THSG）的毒性并比较其差异。方法 2D和3D HepaRG细胞模型给予不同浓度的PMEE和THSG处理24、48、144 h后,检测细胞存活率、DNA损伤程度和细胞凋亡。结果 PMEE可明显抑制2D模型中HepaRG的增殖,但对3D模型中HepaRG的抑制作用不明显;PMEE对2D模型致DNA损伤作用也强于3D模型。THSG对3D模型中HepaRG的抑制作用强于2D模型,且在3D模型的碱性彗星试验中,THSG高剂量组的凋亡细胞发生率显著升高（P<0.01）。结论 3D肝培养模型较2D肝培养模型灵敏度高,PMEE与THSG均具有一定的肝脏毒性,经3D肝培养模型代谢后,PMEE的毒性有所减弱,THSG的毒性则有所增强。

关键词: 何首乌, 二苯乙烯苷, 肝毒性, 二维培养, 三维培养

Abstract: Objective To evaluate the hepatotoxicity of PMEE and THSG on 2D and 3D hepatocyte models in vitro and compare the differences between them. Methods 2D and 3D hepatocyte models were dealt with different concentrations of PMEE and THSG for 24 h, 48 h and 144 h, then the cell viability, DNA damage and cell apoptosis were detected. Results PMEE could inhibit the proliferation of HepaRG in 2D model notably, but the effect on 3D model was not like this. Compared with 3D model, PMEE also had a stronger DNA damage effect on 2D model. The inhibitory effect of THSG on HepaRG in 3D model was greater than that in 2D model. What' s more, the frequency of apoptosis cells of high-dose THSG group was increased notably compared with control group in the alkaline comet assay of 3D model (P <0.01). Conclusion 3D model was greater than 2D model in sensitivity. Both PMEE and THSG had hepatotoxicity and the hepatotoxicity of PMEE was attenuated after being metabolized by 3D hepatocyte models, THSG was the opposite.

Key words: polygonum multiflorum, stilbene glycoside, hepatotoxicity, two-dimension culture, three-dimension culture

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