环境中可致肝脏损伤化学成分体外筛选模型的建立

高美琪; 林鹤; 于娜; 刘兆泉; 任利翔

doi:10.7524/AJE.1673-5897.20191024001

生态毒理学报

, , , ,

高美琪, 林鹤, 于娜, 刘兆泉, 任利翔. 环境中可致肝脏损伤化学成分体外筛选模型的建立[J]. 生态毒理学报, 2020, 15(3): 56-63. doi: 10.7524/AJE.1673-5897.20191024001

引用本文:

高美琪, 林鹤, 于娜, 刘兆泉, 任利翔. 环境中可致肝脏损伤化学成分体外筛选模型的建立[J]. 生态毒理学报, 2020, 15(3): 56-63. doi: 10.7524/AJE.1673-5897.20191024001

Gao Meiqi, Lin He, Yu Na, Liu Zhaoquan, Ren Lixiang. Establishment of an in vitro Screening Model for Environmental Chemicals that Can Induce Liver Damage[J]. Asian Journal of Ecotoxicology, 2020, 15(3): 56-63. doi: 10.7524/AJE.1673-5897.20191024001

Citation:

Gao Meiqi, Lin He, Yu Na, Liu Zhaoquan, Ren Lixiang. Establishment of an in vitro Screening Model for Environmental Chemicals that Can Induce Liver Damage[J]. Asian Journal of Ecotoxicology, 2020, 15(3): 56-63. doi: 10.7524/AJE.1673-5897.20191024001

环境中可致肝脏损伤化学成分体外筛选模型的建立

1. 沈阳化工研究院有限公司安全评价中心, 沈阳 110021;
2. 沈阳药科大学生命科学与生物制药学院, 沈阳 110016

作者简介: 高美琪(1994-),女,硕士,研究方向为肿瘤药理学、替代毒理学,E-mail:1014773401@qq.com

中图分类号: X171.5

Establishment of an in vitro Screening Model for Environmental Chemicals that Can Induce Liver Damage

1. Shenyang Research Institute of Chemical Industry, Safety Evaluation Center, Shenyang 110021, China;
2. Faculty of Life Sciences and Biological Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China

摘要: 环境污染日益严重，环境中的有害化学成分对人类的身体健康具有潜在的危害，而肝脏作为代谢的主要器官，无疑最易受到损伤。本研究旨在建立一种高效的、多参数的肝脏毒性体外筛选方法。采用了5种肝毒性阳性化合物和3种阴性化合物，3种试验方法对2种肝细胞进行了比较。采用四甲基偶氮唑盐微量酶反应比色法（MTT法）考察了化合物对2种细胞体外的生长抑制作用，采用高内涵检测法考察了化合物对肝损伤相关指标的影响，采用实时定量聚合酶链式反应（PCR法）比较了2种细胞的肝药酶含量。5种明确能够导致肝损伤的阳性化学物质对HepG2和L-02细胞均有不同程度的体外生长抑制作用，而3种明确不能诱导肝损伤的化合物对2种细胞均无明显生长抑制作用，且L-02细胞对肝毒性阳性化学物相对敏感。高内涵检测结果表明，不同毒性机制的阳性化合物作用于2种细胞时，与肝损伤机制相关的参数呈现出不同程度的变化，且L-02细胞预测肝损伤的精确度、灵敏度和特异度更高，表明其预测肝损伤的能力更强。此外，L-02细胞药物代谢酶的mRNA表达含量显著高于HepG2细胞，表明其与体内肝脏代谢水平更加接近。综上所述，L-02细胞结合高内涵检测为合适的肝脏毒性体外筛选模型。
- 肝细胞 /
- 肝脏毒性 /
- 毒性筛选 /
- 高内涵检测
Abstract: The environmental pollution pose a threat to human health when exposed to harmful chemicals. Liver, mainly for metabolism, is undoubtedly the most sensitive organ to those harmful chemicals. In the present study, we established an effective in vitro screening model with multiple parameters for evaluation of hepatotoxicity. Five hepatotoxic chemicals and a plurality of three non-hepatotoxic chemicals were selected as positive controls and negative controls, respectively. Meanwhile, three kinds of assay were also applied in two hepatocytes for comparison. The growth inhibitions in these cell types induced by these chemicals were examined using MTT assay. Close relevant indicators of liver injury were then checked using high-content detection method. Hepatic enzyme contents were finally tested by real-time quantitative PCR. Five hepatotoxic chemicals showed significant inhibition on the growth of HepG2 and L-02 cells in different patterns. Furthermore, when those hepatotoxic chemicals were administrated in these cells, the liver injury-related parameters exhibited distinct degrees of changes. Our finding indicates that L-02 cells showed obvious higher sensitivity and specificity than HepG2 cells in hepatotoxic evaluation. The mRNA levels of metabolizing enzymes were also remarkably higher in L-02 cells than in HepG2 cells, indicating that L-02 cell is suitable for in vitro model closely simulating liver metabolism levels in vivo for hepatotoxic screening.
- hepatocytes /
- hepatotoxic /
- toxicity screening /
- high-content detection

Visentin M, Lenggenhager D, Gai Z, et al. Drug-induced bile duct injury[J]. Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease, 2018, 1864:1498-1506

Woodhead J L, Brock W J, Roth S E, et al. Application of a mechanistic model to evaluate putative mechanisms of tolvaptan drug-induced liver injury and identify patient susceptibility factors[J]. Toxicological Sciences, 2017, 155(1):61-74

Hamilton L A, Collins Y A, Collins R E. Drug-induced liver injury[J]. Advanced Critical Care Nursing, 2016, 27(4):430-440

Zhang J, Doshi U, Suzuki A, et al. Evaluation of multiple mechanism-based toxicity endpoints in primary cultured human hepatocytes for the identification of drugs with clinical hepatotoxicity:Results from 152 marketed drugs with known liver injury profiles[J]. Chemico-Biological Interactions, 2016, 255:3-11

Ye H, Nelson L J, Gómez D M M, et al. Dissecting the molecular pathophysiology of drug-induced liver injury[J]. World Journal of Gastroenterology, 2018, 24:1373-1385

Donato M T, Gómez-Lechón M J, Tolosa L. Using high-content screening technology for studying drug-induced hepatotoxicity in preclinical studies[J]. Expert Opinion on Drug Discovery, 2017, 12(2):201-211

Mandavilli B S, Aggeler R J, Chambers K M. Tools to measure cell health and cytotoxicity using high content imaging and analysis[J]. Methods in Molecular Biology, 2018, 1683:33-46

Tolosa L, Gómez-L M J, Donato M T. High-content screening technology for studying drug-induced hepatotoxicity in cell models[J]. Archives of Toxicology, 2015, 89(7):1007-1022

Zhang J, Wang S, Xu L, et al. Multiple perspectives of qingkailing injection-fraction-single compound in revealing the hepatotoxicity of baicalin and hyodeoxycholic acid[J]. Journal of Ethnopharmacology, 2017, 215:147-155

Wu Y, Geng X C, Wang J F, et al. The HepaRG cell line, a superior in vitro model to L-02, HepG2 and hiHeps cell lines for assessing drug-induced liver injury[J]. Cell Biology and Toxicology, 2016, 32(1):37-59

Thakkar S, Tong W, Chen M, et al. The Liver Toxicity Knowledge Base (LKTB) and drug-induced liver injury (DILI) classification for assessment of human liver injury[J]. Expert Review of Gastroenterology & Hepatology, 2017, 12(1):31-38

Yu K, Zhang J, Chen M, et al. Mining hidden knowledge for drug safety assessment:Topic modeling of LiverTox as a case study[J]. BMC Bioinformatics, 2014, 15(Suppl 17):S6

Li X, Li X Y, Huang N, et al. A comprehensive review and perspectives on pharmacology and toxicology of saikosaponins[J]. Phytomedicine, 2018, 50:73-87

Moo L B, Cheul L W, Young J J, et al. Clinical features of drug-induced liver injury according to etiology[J]. Journal of Korean Medical Science, 2015, 30(12):1815-1820

点击查看大图

计量

文章访问数: 2370
HTML全文浏览数: 2370
PDF下载数: 121
施引文献: 0

高美琪, 林鹤, 于娜, 刘兆泉, 任利翔. 环境中可致肝脏损伤化学成分体外筛选模型的建立[J]. 生态毒理学报, 2020, 15(3): 56-63. doi: 10.7524/AJE.1673-5897.20191024001

引用本文:

高美琪, 林鹤, 于娜, 刘兆泉, 任利翔. 环境中可致肝脏损伤化学成分体外筛选模型的建立[J]. 生态毒理学报, 2020, 15(3): 56-63. doi: 10.7524/AJE.1673-5897.20191024001

Citation:

环境中可致肝脏损伤化学成分体外筛选模型的建立

作者简介: 高美琪(1994-),女,硕士,研究方向为肿瘤药理学、替代毒理学,E-mail:1014773401@qq.com
1. 沈阳化工研究院有限公司安全评价中心, 沈阳 110021;

2. 沈阳药科大学生命科学与生物制药学院, 沈阳 110016

收稿日期: 2019-10-24

关键词:

摘要: 环境污染日益严重，环境中的有害化学成分对人类的身体健康具有潜在的危害，而肝脏作为代谢的主要器官，无疑最易受到损伤。本研究旨在建立一种高效的、多参数的肝脏毒性体外筛选方法。采用了5种肝毒性阳性化合物和3种阴性化合物，3种试验方法对2种肝细胞进行了比较。采用四甲基偶氮唑盐微量酶反应比色法（MTT法）考察了化合物对2种细胞体外的生长抑制作用，采用高内涵检测法考察了化合物对肝损伤相关指标的影响，采用实时定量聚合酶链式反应（PCR法）比较了2种细胞的肝药酶含量。5种明确能够导致肝损伤的阳性化学物质对HepG2和L-02细胞均有不同程度的体外生长抑制作用，而3种明确不能诱导肝损伤的化合物对2种细胞均无明显生长抑制作用，且L-02细胞对肝毒性阳性化学物相对敏感。高内涵检测结果表明，不同毒性机制的阳性化合物作用于2种细胞时，与肝损伤机制相关的参数呈现出不同程度的变化，且L-02细胞预测肝损伤的精确度、灵敏度和特异度更高，表明其预测肝损伤的能力更强。此外，L-02细胞药物代谢酶的mRNA表达含量显著高于HepG2细胞，表明其与体内肝脏代谢水平更加接近。综上所述，L-02细胞结合高内涵检测为合适的肝脏毒性体外筛选模型。

English Abstract

Establishment of an in vitro Screening Model for Environmental Chemicals that Can Induce Liver Damage

1. Shenyang Research Institute of Chemical Industry, Safety Evaluation Center, Shenyang 110021, China;
2. Faculty of Life Sciences and Biological Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China

Received Date: 2019-10-24

Keywords:

Abstract: The environmental pollution pose a threat to human health when exposed to harmful chemicals. Liver, mainly for metabolism, is undoubtedly the most sensitive organ to those harmful chemicals. In the present study, we established an effective in vitro screening model with multiple parameters for evaluation of hepatotoxicity. Five hepatotoxic chemicals and a plurality of three non-hepatotoxic chemicals were selected as positive controls and negative controls, respectively. Meanwhile, three kinds of assay were also applied in two hepatocytes for comparison. The growth inhibitions in these cell types induced by these chemicals were examined using MTT assay. Close relevant indicators of liver injury were then checked using high-content detection method. Hepatic enzyme contents were finally tested by real-time quantitative PCR. Five hepatotoxic chemicals showed significant inhibition on the growth of HepG2 and L-02 cells in different patterns. Furthermore, when those hepatotoxic chemicals were administrated in these cells, the liver injury-related parameters exhibited distinct degrees of changes. Our finding indicates that L-02 cells showed obvious higher sensitivity and specificity than HepG2 cells in hepatotoxic evaluation. The mRNA levels of metabolizing enzymes were also remarkably higher in L-02 cells than in HepG2 cells, indicating that L-02 cell is suitable for in vitro model closely simulating liver metabolism levels in vivo for hepatotoxic screening.

全文HTML

参考文献 (14)

下载: 全尺寸图片幻灯片

返回文章

用微信扫码二维码

分享至好友和朋友圈

环境中可致肝脏损伤化学成分体外筛选模型的建立

作者简介: 高美琪(1994-),女,硕士,研究方向为肿瘤药理学、替代毒理学,E-mail:1014773401@qq.com

Establishment of an in vitro Screening Model for Environmental Chemicals that Can Induce Liver Damage

计量

出版历程

环境中可致肝脏损伤化学成分体外筛选模型的建立

作者简介: 高美琪(1994-),女,硕士,研究方向为肿瘤药理学、替代毒理学,E-mail:1014773401@qq.com 1. 沈阳化工研究院有限公司安全评价中心, 沈阳 110021; 2. 沈阳药科大学生命科学与生物制药学院, 沈阳 110016

English Abstract

Establishment of an in vitro Screening Model for Environmental Chemicals that Can Induce Liver Damage

全文HTML

目录

作者简介: 高美琪(1994-),女,硕士,研究方向为肿瘤药理学、替代毒理学,E-mail:1014773401@qq.com
1. 沈阳化工研究院有限公司安全评价中心, 沈阳 110021;

2. 沈阳药科大学生命科学与生物制药学院, 沈阳 110016