A novel strategy for identifying hepatotoxic constituents in traditional Chinese medicine using dose-normalized intracellular accumulation as a cytotoxicity indicator: a case study of Jinlingzi San

IntroductionHerb-induced liver injury associated with traditional Chinese medicine (TCM) has increasingly attracted scientific attention. However, the rapid and effective methods for elucidating the material basis of hepatotoxicity in TCM are still lacking. This study developed a strategy based on t...

Full description

Saved in:
Bibliographic Details
Main Authors: Xiaoting Gu, Hanyang Wang, Keran Li, Cuiting Wu, Xin Di
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Pharmacology
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/fphar.2025.1585186/full
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850122471033798656
author Xiaoting Gu
Xiaoting Gu
Hanyang Wang
Hanyang Wang
Keran Li
Cuiting Wu
Xin Di
author_facet Xiaoting Gu
Xiaoting Gu
Hanyang Wang
Hanyang Wang
Keran Li
Cuiting Wu
Xin Di
author_sort Xiaoting Gu
collection DOAJ
description IntroductionHerb-induced liver injury associated with traditional Chinese medicine (TCM) has increasingly attracted scientific attention. However, the rapid and effective methods for elucidating the material basis of hepatotoxicity in TCM are still lacking. This study developed a strategy based on the use of dose-normalized intracellular accumulation as a cytotoxicity indicator to identify key hepatotoxic components in TCM.MethodsJinlingzi San (JLZS) composed of Fructus Toosendan (FT) and Rhizoma Corydalis (RC) was used as a model sample in this study. Hepatotoxicity was evaluated through both in vivo (14-day continuous gavage administration in rats) and in vitro (24-hour co-incubation with L-02 cells) models. Chemical components in JLZS extract and those accumulated in L-02 cells were identified using LC-MS/MS. The intracellular accumulations of multiple components after exposing L-02 cells to the extracts of JLZS, FT and RC were determined.ResultsJLZS administration resulted in subacute liver injury in rats and demonstrated cytotoxicity to L-02 cells. Seven analytes (coptisine, tetrahydrocoptisine, berberine, jatrorrhizine, palmatine, dehydrocorydaline and toosendanin) were identified in cell lysates following incubation with JLZS. Coptisine and berberine were regarded as the main potential hepatotoxic components in JLZS for its highest dose-normalized intracellular accumulation and the strongest cytotoxicity. In addition, the intracellular accumulations of coptisine and berberine were lower in the JLZS group compared to the RC group alone.DiscussionThe findings suggest that dose-normalized intracellular accumulation serves as a reliable indicator for identifying hepatotoxic constituents in TCM. The observed reduction in intracellular accumulation of coptisine and berberine in the JLZS formulation compared to RC alone maybe reflect the scientific meaning of detoxicity by compatibility with FT and RC.
format Article
id doaj-art-4ce41aaa5217481487280883bb3f0eba
institution OA Journals
issn 1663-9812
language English
publishDate 2025-05-01
publisher Frontiers Media S.A.
record_format Article
series Frontiers in Pharmacology
spelling doaj-art-4ce41aaa5217481487280883bb3f0eba2025-08-20T02:34:50ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122025-05-011610.3389/fphar.2025.15851861585186A novel strategy for identifying hepatotoxic constituents in traditional Chinese medicine using dose-normalized intracellular accumulation as a cytotoxicity indicator: a case study of Jinlingzi SanXiaoting Gu0Xiaoting Gu1Hanyang Wang2Hanyang Wang3Keran Li4Cuiting Wu5Xin Di6State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, ChinaLaboratory of Drug Metabolism and Pharmacokinetics, School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, Liaoning, ChinaLaboratory of Drug Metabolism and Pharmacokinetics, School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, Liaoning, ChinaCentre for Applied Pharmacokinetic Research, The University of Manchester, Manchester, United KingdomState Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, ChinaLaboratory of Drug Metabolism and Pharmacokinetics, School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, Liaoning, ChinaLaboratory of Drug Metabolism and Pharmacokinetics, School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, Liaoning, ChinaIntroductionHerb-induced liver injury associated with traditional Chinese medicine (TCM) has increasingly attracted scientific attention. However, the rapid and effective methods for elucidating the material basis of hepatotoxicity in TCM are still lacking. This study developed a strategy based on the use of dose-normalized intracellular accumulation as a cytotoxicity indicator to identify key hepatotoxic components in TCM.MethodsJinlingzi San (JLZS) composed of Fructus Toosendan (FT) and Rhizoma Corydalis (RC) was used as a model sample in this study. Hepatotoxicity was evaluated through both in vivo (14-day continuous gavage administration in rats) and in vitro (24-hour co-incubation with L-02 cells) models. Chemical components in JLZS extract and those accumulated in L-02 cells were identified using LC-MS/MS. The intracellular accumulations of multiple components after exposing L-02 cells to the extracts of JLZS, FT and RC were determined.ResultsJLZS administration resulted in subacute liver injury in rats and demonstrated cytotoxicity to L-02 cells. Seven analytes (coptisine, tetrahydrocoptisine, berberine, jatrorrhizine, palmatine, dehydrocorydaline and toosendanin) were identified in cell lysates following incubation with JLZS. Coptisine and berberine were regarded as the main potential hepatotoxic components in JLZS for its highest dose-normalized intracellular accumulation and the strongest cytotoxicity. In addition, the intracellular accumulations of coptisine and berberine were lower in the JLZS group compared to the RC group alone.DiscussionThe findings suggest that dose-normalized intracellular accumulation serves as a reliable indicator for identifying hepatotoxic constituents in TCM. The observed reduction in intracellular accumulation of coptisine and berberine in the JLZS formulation compared to RC alone maybe reflect the scientific meaning of detoxicity by compatibility with FT and RC.https://www.frontiersin.org/articles/10.3389/fphar.2025.1585186/fulltraditional Chinese medicineshepatotoxicitydose-normalized intracellular accumulationLC-MS/MSJinlingzi San
spellingShingle Xiaoting Gu
Xiaoting Gu
Hanyang Wang
Hanyang Wang
Keran Li
Cuiting Wu
Xin Di
A novel strategy for identifying hepatotoxic constituents in traditional Chinese medicine using dose-normalized intracellular accumulation as a cytotoxicity indicator: a case study of Jinlingzi San
Frontiers in Pharmacology
traditional Chinese medicines
hepatotoxicity
dose-normalized intracellular accumulation
LC-MS/MS
Jinlingzi San
title A novel strategy for identifying hepatotoxic constituents in traditional Chinese medicine using dose-normalized intracellular accumulation as a cytotoxicity indicator: a case study of Jinlingzi San
title_full A novel strategy for identifying hepatotoxic constituents in traditional Chinese medicine using dose-normalized intracellular accumulation as a cytotoxicity indicator: a case study of Jinlingzi San
title_fullStr A novel strategy for identifying hepatotoxic constituents in traditional Chinese medicine using dose-normalized intracellular accumulation as a cytotoxicity indicator: a case study of Jinlingzi San
title_full_unstemmed A novel strategy for identifying hepatotoxic constituents in traditional Chinese medicine using dose-normalized intracellular accumulation as a cytotoxicity indicator: a case study of Jinlingzi San
title_short A novel strategy for identifying hepatotoxic constituents in traditional Chinese medicine using dose-normalized intracellular accumulation as a cytotoxicity indicator: a case study of Jinlingzi San
title_sort novel strategy for identifying hepatotoxic constituents in traditional chinese medicine using dose normalized intracellular accumulation as a cytotoxicity indicator a case study of jinlingzi san
topic traditional Chinese medicines
hepatotoxicity
dose-normalized intracellular accumulation
LC-MS/MS
Jinlingzi San
url https://www.frontiersin.org/articles/10.3389/fphar.2025.1585186/full
work_keys_str_mv AT xiaotinggu anovelstrategyforidentifyinghepatotoxicconstituentsintraditionalchinesemedicineusingdosenormalizedintracellularaccumulationasacytotoxicityindicatoracasestudyofjinlingzisan
AT xiaotinggu anovelstrategyforidentifyinghepatotoxicconstituentsintraditionalchinesemedicineusingdosenormalizedintracellularaccumulationasacytotoxicityindicatoracasestudyofjinlingzisan
AT hanyangwang anovelstrategyforidentifyinghepatotoxicconstituentsintraditionalchinesemedicineusingdosenormalizedintracellularaccumulationasacytotoxicityindicatoracasestudyofjinlingzisan
AT hanyangwang anovelstrategyforidentifyinghepatotoxicconstituentsintraditionalchinesemedicineusingdosenormalizedintracellularaccumulationasacytotoxicityindicatoracasestudyofjinlingzisan
AT keranli anovelstrategyforidentifyinghepatotoxicconstituentsintraditionalchinesemedicineusingdosenormalizedintracellularaccumulationasacytotoxicityindicatoracasestudyofjinlingzisan
AT cuitingwu anovelstrategyforidentifyinghepatotoxicconstituentsintraditionalchinesemedicineusingdosenormalizedintracellularaccumulationasacytotoxicityindicatoracasestudyofjinlingzisan
AT xindi anovelstrategyforidentifyinghepatotoxicconstituentsintraditionalchinesemedicineusingdosenormalizedintracellularaccumulationasacytotoxicityindicatoracasestudyofjinlingzisan
AT xiaotinggu novelstrategyforidentifyinghepatotoxicconstituentsintraditionalchinesemedicineusingdosenormalizedintracellularaccumulationasacytotoxicityindicatoracasestudyofjinlingzisan
AT xiaotinggu novelstrategyforidentifyinghepatotoxicconstituentsintraditionalchinesemedicineusingdosenormalizedintracellularaccumulationasacytotoxicityindicatoracasestudyofjinlingzisan
AT hanyangwang novelstrategyforidentifyinghepatotoxicconstituentsintraditionalchinesemedicineusingdosenormalizedintracellularaccumulationasacytotoxicityindicatoracasestudyofjinlingzisan
AT hanyangwang novelstrategyforidentifyinghepatotoxicconstituentsintraditionalchinesemedicineusingdosenormalizedintracellularaccumulationasacytotoxicityindicatoracasestudyofjinlingzisan
AT keranli novelstrategyforidentifyinghepatotoxicconstituentsintraditionalchinesemedicineusingdosenormalizedintracellularaccumulationasacytotoxicityindicatoracasestudyofjinlingzisan
AT cuitingwu novelstrategyforidentifyinghepatotoxicconstituentsintraditionalchinesemedicineusingdosenormalizedintracellularaccumulationasacytotoxicityindicatoracasestudyofjinlingzisan
AT xindi novelstrategyforidentifyinghepatotoxicconstituentsintraditionalchinesemedicineusingdosenormalizedintracellularaccumulationasacytotoxicityindicatoracasestudyofjinlingzisan