phenanthrenes and Chemical-and-Drug-Induced-Liver-Injury

Triptolide is a predominant active component of Triptergium wilfordii Hook. F, which has been used for the treatment of cancers and autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus and diabetic nephropathy. Therefore, triptolide and its derivates are considered to have promising prospects for development into drugs. However, the clinical application of triptolide is limited due to various organ toxicities, especially liver toxicity. The potential mechanism of triptolide-induced hepatotoxicity has attracted increasing attention. Over the past five years, studies have revealed that triptolide-induced liver toxicity is involved in metabolic imbalance, oxidative stress, inflammations, autophagy, apoptosis, and the regulation of cytochrome P450 (CYP450) enzymes, gut microbiota and immune cells. In this review, we summarize the pharmacological applications and hepatotoxicity mechanism of triptolide, which will provide solid theoretical evidence for further research of triptolide.

Topics: Chemical and Drug Induced Liver Injury; Diterpenes; Drug-Related Side Effects and Adverse Reactions; Epoxy Compounds; Hepatitis; Humans; Phenanthrenes

This study was to investigate the potential mechanism of triptolide-induced hepatotoxicity. We found a novel and variable role of p53/Nrf2 crosstalk in triptolide-induced hepatotoxic process. Low doses of triptolide led to adaptive stress response without obvious toxicity, while high levels of triptolide caused severe adversity. Correspondingly, at the lower levels of triptolide treatment, nuclear translocation of Nrf2 as well as its downstream efflux transporters multidrug resistance proteins and bile salt export pump expressions were significantly enhanced, so did p53 pathways that also increased; at a toxic concentration, total and nuclear accumulations of Nrf2 decreased, while p53 showed an obvious nuclear translocation. Further studies showed the cross-regulation between p53 and Nrf2 after different concentrations of triptolide treatment. Under mild stress conditions, Nrf2 induced p53 highly expression to maintain the pro-survival outcome, while p53 showed no obvious effect on Nrf2 expression and transcriptional activity. Under high stress conditions, the remaining Nrf2 as well as the largely induced p53 mutually inhibited each other, leading to a hepatotoxic result. Nrf2 and p53 could physically and dynamically interact. Low levels of triptolide enhanced the interaction between Nrf2 and p53. Reversely, p53/Nrf2 complex dissociated at high levels of triptolide treatment. Altogether, variable p53/Nrf2 crosstalk contributes to triptolide-induced self-protection and hepatotoxicity, modulation of which may be a potential strategy for triptolide-induced hepatotoxicity intervention.

Topics: Chemical and Drug Induced Liver Injury; Diterpenes; Drug-Related Side Effects and Adverse Reactions; Epoxy Compounds; Humans; NF-E2-Related Factor 2; Phenanthrenes; Tumor Suppressor Protein p53

Liver toxicity induced by Triptolide (TP) has limited its clinical application on rheumatoid arthritis (RA). Saponins have been proved as an efficacious remedy to mitigate hepatotoxicity. However, the mechanism of reducing hepatotoxicity by saponins intervention remains incompletely characterized. Tryptophan (Trp) metabolites activate transcriptional regulators to mediate host detoxification responses. Our study aimed to investigate whether Clematichinenoside AR (C-AR) could attenuate TP-induced liver damage by regulating Trp metabolism. We used targeted metabolomics to quantify Trp metabolites in the serum and liver samples of collagen-induced arthritis rats treated by TP. Multiple comparison analyses helped the evaluation of promising biomarkers. The pronounced changed levels of Trp, indole acetic acid, and indole-3-carboxaldehyde in the serum and indole acetic acid, indole, and tryptamine in the liver are relevant to TP-induced liver injury. Intervention with C-AR could relieve TP-induced hepatotoxicity evidenced by ameliorative serum parameters and hepatic histology. In addition, C-AR regulated the levels of these indoles biomarker candidates to normal. Therapeutic modulation with natural compounds might be a useful clinical strategy to ameliorate toxicity induced by TP. Deciphering Trp metabolism will facilitate a better understanding of the pathogenesis of diseases and drug responding.

Topics: Animals; Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Liver; Phenanthrenes; Rats; Saponins; Triterpenes; Tryptophan

Tripterygium glycosides tablets (TGT) and Tripterygium wilfordii tablets (TWT) have been used to treat autoimmune diseases clinically, however, the side effects of TWT are higher than TGT, especially for hepatotoxicity.. This study aims to determine the mechanism of TWT-induced liver injury.. We performed metabolomic analysis of samples from mice with liver injury induced by TGT and TWT. Ppara-null mice were used to determine the role of PPARα in TWT-induced liver injury.. The results indicated that TWT induced the accumulation of medium- and long-chain carnitines metabolism, which was associated with the disruption of PPARα-IL6-STAT3 axis. PPARα agonists fenofibrate could reverse the liver injury from TWT and TP/Cel, and its protective role could be attenuated in Ppara-null mice. The toxicity difference of TWT and TGT was due to the different ratio of triptolide (TP) and celastrol (Cel) in the tablet in which TP/Cel was lower in TWT than TGT. The hepatotoxicity induced by TP and Cel also inhibited PPARα and upregulated IL6-STAT3 axis, which could be alleviated following by PPARα activation.. These results indicated that PPARα plays an important role in the hepatotoxicity of Tripterygium wilfordii, and PPARα activation may offer a promising approach to prevent hepatotoxicity induced by the preparations of Tripterygium wilfordii.

Topics: Animals; Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Male; Metabolomics; Mice; Mice, Inbred C57BL; Mice, Knockout; Pentacyclic Triterpenes; Phenanthrenes; Plant Extracts; PPAR alpha; Tablets; Tripterygium

Triptolide (TP) exhibits extensive pharmacological activity, but its hepatotoxicity and intestinal injury are significant and limit its clinical use.. To investigate the effect of gut microbiota disturbance after antibiotic pretreatment on TP-induced hepatotoxicity, intestinal injury and their mechanism.. We compared the characteristics of TP-induced hepatotoxicity and intestinal injury in mice with or without antibiotic pretreatment. The levels of cytokines in the serum, immunohistochemistry, and the pharmacokinetics of TP were determined.. Antibiotic pretreatment aggravates TP-induced hepatotoxicity and ileum/colon injury. TP induces hepatotoxicity in a dose-dependent manner after antibiotic pretreatment. Serum IL-1β and IL-6 levels were increased in mice given oral TP after antibiotic pretreatment. TP can increase the expression of NLRP3 inflammasome in hepatocytes, and Oral TP after antibiotic pretreatment can significantly enhance its expression, but NLRP3 inflammasome no significant change in colon and ileum. The pharmacokinetic characteristics of TP are altered significantly by antibiotic pretreatment, as shown by a 145.87% increase in C. Antibiotic pretreatment aggravates triptolide-induced hepatotoxicity and intestinal injury through elevated inflammatory response and promoted triptolide absorption.

Topics: Animals; Anti-Bacterial Agents; Chemical and Drug Induced Liver Injury; Diterpenes; Drug-Related Side Effects and Adverse Reactions; Epoxy Compounds; Inflammasomes; Liver; Mice; NLR Family, Pyrin Domain-Containing 3 Protein; Phenanthrenes

Triptolide (TP) is a major active ingredient and toxic component of Tripterygium wilfordii Hook F (TWHF), which exhibits multiple activities and remarkable hepatotoxicity, the latter of which limits its clinical application due to the risk of liver injury. Previous research has revealed the hepatotoxicity of TP resulting in liver hypersensitivity upon lipopolysaccharide (LPS) stimulation. However, existing research has not elucidated the potential immune mechanism such as Th17/Treg imbalance in TP-induced hepatic excessive immune response to exogenous LPS.. To investigate the role of Th17/Treg imbalance in TP-induced hepatic excessive immune response to exogenous LPS.. Mice were administered with TP, LPS, neutralization antibody and small molecule inhibitor respectively. Serum transaminase level was measured to determine the severity of liver injury. Frequencies of liver Th17 and Treg cells were analyzed by flow cytometry. Serum cytokine levels were performed by ELSIA, and mRNA levels of liver cytokine were performed by qPCR. The status of neutrophil infiltration was performed by myeloperoxidase (MPO) IHC measurement. Morphological observation of liver was performed by hematoxylin and eosin (H&E) staining.. Mice given a single intragastric dose of TP (500 μg/kg) developed lethal fulminant hepatitis following intraperitoneal injection of LPS (0.1 mg/kg), characterized by low survival rate, severe liver injury, high levels of inflammation and neutrophil infiltration. Hepatic Th17/Treg imbalance emerged together with liver injury in these mice. Neutralization of IL-17A attenuated the liver injury and ameliorated the neutrophil infiltration. The TP-induced alteration of hepatic Th17/Treg balance was closely related to the outcome of immune-mediated acute liver injury triggered by LPS. Pretreatment with the STAT3 inhibitor AG490 effectively restored Th17/Treg balance, significantly reducing the production of IL-17A and finally attenuating the degree of liver injury.. Hepatic Th17/Treg imbalance not only exacerbates TP- and LPS-induced liver injury, but also serves as an indispensable part in the mechanisms of TP-induced hepatic intolerance to exogenous endotoxin.

Topics: Animals; Chemical and Drug Induced Liver Injury; Cytokines; Diterpenes; Epoxy Compounds; Immunity; Interleukin-17; Lipopolysaccharides; Mice; Mice, Inbred C57BL; Phenanthrenes; T-Lymphocytes, Regulatory; Th17 Cells

Triptolide (TP) has limited usage in clinical practice due to its side effects and toxicity, especially liver injury. Hepatic macrophages, key player of liver innate immunity, were found to be recruited and activated by TP in our previous study. The nuclear factor-erythroid-2-related factor 2 (Nrf2) pathway exerts a protective role in TP-induced liver damage, but its effect on the functions of hepatic macrophage has not been elucidated. Here, we determined whether TP can regulate the recruitment and polarization of hepatic macrophages by inhibiting Nrf2 signaling cascade. Our results demonstrated that TP inhibited the Nrf2 signaling pathway in hepatic macrophages. The changes in hepatic macrophages were responsible for the increased susceptibility toward inflammatory stimuli, and hence, TP pretreatment could induce severe liver damage upon the stimulation of a nontoxic dose of lipopolysaccharides. In addition, the Nrf2 agonist protected macrophages from TP-induced toxicity and Nrf2 deficiency significantly aggravated liver injury by enhancing the recruitment and M1 polarization of hepatic macrophages. This study suggests that Nrf2 pathway-mediated hepatic macrophage polarization plays an essential role in TP-induced liver damage, which can serve as a potential therapeutic target for preventing hepatotoxicity induced by TP.

Topics: Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Humans; Liver; Macrophages; NF-E2-Related Factor 2; Phenanthrenes; Signal Transduction

The liver is an important organ for amino acid metabolism, and its damage can be reflected in the changes of amino acid level in the body. Triptolide (TP) has broad anti-inflammatory and anti-tumor activities, but its clinical application is limited due to hepatotoxicity. In this work, a simple, accurate and sensitive gas chromatography-triple quadrupole mass spectrometry (GC-QqQ-MS/MS) method was developed and validated for evaluating the serum levels of amino acids from control and TP-induced liver injured rats, and chemometric analysis was employed for amino acid metabolic profiles analysis. It was found that 11 amino acids showed significant changes after TP administration, and they were mainly involved in 5 metabolic pathways that are phenylalanine, tyrosine and tryptophan biosynthesis, alanine, aspartate and glutamate metabolism, glutamine and glutamate metabolism, phenylalanine metabolism and arginine biosynthesis. Five amino acids including tyrosine, glutamine, glutamic acid, tryptophan and alanine were identified as biomarkers of TP hepatotoxicity by further analysis. These results indicated that the novel amino acid metabolic profiling study based on the GC-QqQ-MS/MS provided not only exact concentrations of serum amino acids, but also a prospective methodology for evaluation of chemically induced liver injury.

Topics: Amino Acids; Animals; Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Female; Gas Chromatography-Mass Spectrometry; Molecular Structure; Phenanthrenes; Rats; Rats, Sprague-Dawley

Triptolide (TP), the main bioactive and toxic ingredient of Tripterygium wilfordii Hook F, causes severe toxicity, particularly for hepatotoxicity. However, the underlying mechanisms for its hepatotoxicity are not entirely clear.. The purpose of the study was to explore the role of miR-155, a microRNA closely related to various liver injuries and a regulator of the nuclear factor erythroid 2-related factor 2 (Nrf2) antioxidant pathway, in TP-induced liver injury in vitro and in vivo.. First, in vitro L02 cells were treated with different concentrations of TP. The protein levels of Nrf2 and its downstream genes Heme oxygenase1 (HO-1) were determined by Western blot. The mRNA expression of miR-155, Nrf2, NAD(P)H: quinone oxidoreductase 1 (NQO1) and HO-1 were measured using qRT-PCR. And we transfected miR-155 inhibitor and miminc before TP treatment to determine the mRNA and/or protein levels of miR-155, Nrf2 and HO-1. Then, we further confirmed the interaction between miR-155 and Nrf2 pathway in TP-induced hepatic injury in BALB/C mice. The degree of liver injury was determined by HE staining and serum biochemical. The mRNA expression of miR-155 was examined with qRT-PCR and Nrf2 and HO-1 gene expression in liver were evaluated by immunohistochemistry and/or Western blot.. The results showed that TP significantly induced the expression of miR-155 both in L02 cells and in rodents liver tissue, and the inhibition of miR-155 could mitigate the hepatic damages caused by TP. Further experiments demonstrated that the inhibition of miR-155 reversed the down-regulation of Nrf2 and HO-1 by TP, while the miR-155 mimic enhanced the effects of TP. Animal experiments also showed that the inhibition of miR-155 by miR-155 antagomir reversed the decrease of Nrf2 induced by TP administration.. These results indicated that miR-155 played an important role in TP-induced hepatotoxicity by regulating the Nrf2 signaling pathway.

Topics: Animals; Cell Line; Cell Survival; Chemical and Drug Induced Liver Injury; Diterpenes; Down-Regulation; Epoxy Compounds; Gene Expression Regulation; Hepatocytes; Humans; Male; Mice; Mice, Inbred BALB C; MicroRNAs; NF-E2-Related Factor 2; Phenanthrenes

How triptolide is associated with mitochondrial dysfunction and apoptosis in connection with its hepatotoxicity remains unclear. The objective of our study was to find out the link between mitochondrial dynamics and cell death in triptolide induced hepatotoxicity. We treated L02 cells with 25 nM concentration of triptolide. The results demonstrated that triptolide treatment caused an increase in apoptotic cell death, mitochondrial depolarization, ROS overproduction, a decrease in ATP production, and mitochondrial fragmentation which in turn is associated with the activation of Drp1 fission protein. Triptolide treatment led to the translocation of Drp1 from the cytosol into outer mitochondrial membrane where it started mitochondrial fission. This fission event is coupled with the mitochondrial release of cytochrome c into the cytosol and subsequently caspase-3 activation. TEM analysis of rat liver tissues revealed the distortion of mitochondrial morphology in triptolide-treated group. Western blot analysis explained that disruption in mitochondrial morphology was attached with the recruitment of Drp1 to mitochondria, cytochrome c release, and caspase-3 activation. However, Mdivi-1 co-treatment inhibited the activation of Drp1 and caspase-3 and blocked the release of cytochrome c into the cytosol. In short, inhibiting Drp1 protein activation may provide a new potential target for curing Drp1-associated apoptosis in triptolide-induced hepatotoxicity.

Topics: Animals; Apoptosis; Cell Line; Cell Survival; Chemical and Drug Induced Liver Injury; Diterpenes; Dose-Response Relationship, Drug; Dynamins; Epoxy Compounds; Female; Hepatocytes; Humans; Mitochondria, Liver; Mitochondrial Dynamics; Phenanthrenes; Rats, Wistar

As a potential drug for treating inflammatory, autoimmune diseases and cancers, triptolide (TP) is greatly limited in clinical practice due to its severe toxicity, particularly for liver injury. Recently, metabolic homeostasis was vitally linked to drug-induced liver injury and gut microbiota was established to play an important role. In this study, we aimed to investigate the functions of gut microbiota on TP-induced hepatotoxicity using metabolomics in mice. Here, predepletion of gut microbiota by antibiotic treatment strikingly aggravated liver injury and caused mortality after treated with a relatively safe dosage of TP at 0.5 mg/kg, which could be reversed by gut microbial transplantation. The loss of gut microbiota prior to TP treatment dramatically elevated long chain fatty acids and bile acids in plasma and liver. Further study suggested that gut microbiota-derived propionate contributed to the protective effect of gut microbiota against TP evidenced by ameliorative inflammatory level (Tnfa, Il6 and Cox2), ATP, malondialdehyde and hepatic histology. Supplementing with propionate significantly decreased the mRNA levels of genes involved in fatty acid biosynthesis (Srebp1c, Fasn and Elovl6), resulting in the decreased long chain fatty acids in liver. Moreover, TP restricted the growth of Firmicutes and led to the deficiency of short chain fatty acids in cecum content. In conclusion, our study warns the risk for TP and its preparations when antibiotics are co-administrated. Intervening by foods, prebiotics and probiotics toward gut microbiota or supplementing with propionate may be a clinical strategy to improve toxicity induced by TP.

Topics: Animals; Anti-Bacterial Agents; Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Fatty Acids, Volatile; Gastrointestinal Microbiome; Liver; Male; Mice, Inbred C57BL; Phenanthrenes; Propionates; Signal Transduction

Triptolide (TP), one of the main bioactive diterpenes of the herbal medicine Tripterygium wilfordii Hook F, is used for the treatment of autoimmune diseases in the clinic and is accompanied by severe hepatotoxicity. CYP3A4 has been reported to be responsible for TP metabolism, but the mechanism remains unclear. The present study applied a UPLC-QTOF-MS-based metabolomics analysis to characterize the effect of CYP3A4 on TP-induced hepatotoxicity. The metabolites carnitines, lysophosphatidylcholines (LPCs) and a serious of amino acids were found to be closely related to liver damage indexes in TP-treated female mice. Metabolomics analysis further revealed that the CYP3A4 inducer dexamethasone improved the level of LPCs and amino acids, and defended against oxidative stress. On the contrary, pretreatment with the CYP3A4 inhibitor ketoconazole increased liver damage with most metabolites being markedly altered, especially carnitines. Among these metabolites, except for LPC18:2, LPC20:1 and arginine, dexamethasone and ketoconazole both affected oxidative stress induced by TP. The current study provides new mechanistic insights into the metabolic alterations, leading to understanding of the role of CYP3A4 in hepatotoxicity induced by TP.

Topics: Animals; Chemical and Drug Induced Liver Injury; Chromatography, High Pressure Liquid; Cytochrome P-450 CYP3A; Cytochrome P-450 CYP3A Inhibitors; Dexamethasone; Diterpenes; Epoxy Compounds; Female; Ketoconazole; Liver; Mass Spectrometry; Metabolome; Metabolomics; Mice; Mice, Inbred C57BL; Phenanthrenes

In general, anti-inflammatory treatment is considered for multiple liver diseases despite the etiology. But current drugs for alleviating liver inflammation have defects, making it necessary to develop more potent and safer drugs for liver injury. In this study, we screened a series of (dihydro-)stilbene or (dihydro-)phenanthrene derivatives extracted from Pholidota chinensis for their potential biological activities. Among 31 compounds, the dihydro-stilbene gigantol exerted most potent protective effects on human hepatocytes against lithocholic acid toxicity, and exhibited solid antioxidative and anti-inflammatory effect in vitro. In mice with CCl

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents; Antioxidants; Bibenzyls; Carbon Tetrachloride; Chemical and Drug Induced Liver Injury; Complement Membrane Attack Complex; Guaiacol; Hepatocytes; Humans; Inflammation; Lipid Peroxidation; Lithocholic Acid; Liver; Male; Mice, Inbred ICR; Oxidative Stress; Phenanthrenes; Proteome; Rats, Sprague-Dawley; Stilbenes

We aimed to determine the diurnal rhythm of Tripterygium wilfordii (TW) hepatotoxicity and to investigate a potential role of metabolism and pharmacokinetics in generating chronotoxicity.. Hepatotoxicity was determined based on assessment of liver injury after dosing mice with TW at different circadian time points. Circadian clock control of metabolism, pharmacokinetics and hepatotoxicity was investigated using Clock-deficient (Clock. Hepatotoxicity of TW displayed a significant circadian rhythm (the highest level of toxicity was observed at ZT2 and the lowest level at ZT14). Pharmacokinetic experiments showed that oral gavage of TW at ZT2 generated higher plasma concentrations (and systemic exposure) of triptolide (a toxic constituent) compared with ZT14 dosing. This was accompanied by reduced formation of triptolide metabolites at ZT2. Loss of Clock gene sensitized mice to TW-induced hepatotoxicity and abolished the time-dependency of toxicity that was well correlated with altered metabolism and pharmacokinetics of triptolide. Loss of Clock gene also decreased Cyp3a11 expression in mouse liver and blunted its diurnal rhythm.. Tripterygium wilfordii chronotoxicity was associated with diurnal variations in triptolide pharmacokinetics and circadian expression of hepatic Cyp3a11 regulated by circadian clock. Our findings may have implications for improving TW treatment outcome with a chronotherapeutic approach.

Topics: Activation, Metabolic; Animals; Chemical and Drug Induced Liver Injury; Circadian Rhythm; CLOCK Proteins; Cytochrome P-450 CYP3A; Diterpenes; Epoxy Compounds; Liver; Male; Membrane Proteins; Mice, Inbred C57BL; Mice, Knockout; Phenanthrenes; Plant Extracts; Toxicokinetics; Tripterygium

Triptolide (TP), as the main component of Tripterygium Wilfordii (TW), can induce obvious liver injury when exerting the therapeutic effect. However, in our previous study, Catalpol (CAT), the main active ingredient of Rehmannia Glutinosa (RG), was shown to increase the drug clearance rate of TP and to attenuate TP-induced hepatotoxicity. Thus the present study aims to address the roles of phase I and II metabolic enzymes and the nuclear receptors in the detoxification process of TP, to analyze the mechanism of CAT reducing hepatotoxicity. For this purpose, SD rats and human liver cell line L-02 and HepG2 cells were selected, and treated with TP or the combination of TP and CAT in our study. Then the effect of CAT on detoxification of TP was analyzed, and the roles of phase I metabolic enzymes cytochrome P450 3A2/4 (CYP3A2/4) and phase II metabolic enzyme UDP-glucuronosyltransferase 1A6 (UGT1A6) and their related nuclear receptor regulations were evaluated. It was found that TP inhibited the transcription of CYP3A2/4. And through the constitutive androstane receptor (CAR) pathway, CAT not only significantly changed this inhibition and increased the expression of CYP3A2/4 but also increased the expression of CYP2C9, both of which are phase I detoxification enzymes of TP. And with the gene-silenced experiment, it was confirmed that this regulation was CAR-dependent. We also found that CAT could continue to exert a certain protective effect after CAR was silenced, with UGT1A6, the phase II detoxification enzyme of TP, significantly induced. And this was closely related to the enhanced transcriptional regulation of the nuclear factor erythroid 2-related factor 2 (NRF2) pathway. In conclusion, our results reveal that CAT can induce TP's phase I detoxification enzymes CYP3A2/4 and CYP2C9 through the CAR pathway, and induce TP's phase II detoxification enzyme UGT1A6 via the NRF2 pathway when CAR is strongly inhibited. And this coordinate regulation of CAT may be an important source of the effect for CAT to increase TP metabolic conversion and reduce TP hepatotoxicity.

Topics: Animals; Chemical and Drug Induced Liver Injury; Constitutive Androstane Receptor; Cytochrome P-450 Enzyme System; Disease Models, Animal; Diterpenes; Epoxy Compounds; Female; Glucuronosyltransferase; Hep G2 Cells; Hepatocytes; Humans; Iridoid Glucosides; Liver; Metabolic Detoxication, Phase I; Metabolic Detoxication, Phase II; NF-E2-Related Factor 2; Phenanthrenes; Rats, Sprague-Dawley; Receptors, Cytoplasmic and Nuclear; Signal Transduction

Triptolide, a major active component of Triptergium wilfordii Hook. f, is used in the treatment of autoimmune disease. However, triptolide is associated with severe adverse reactions, especially hepatotoxicity, which limits its clinical application. To examine the underlying mechanism of triptolide-induced liver injury, a combination of dose- and time-dependent toxic effects, RNA-seq and metabolomics were employed. Triptolide-induced toxicity occurred in a dose- and time-dependent manners and was characterized by apoptosis and not necroptosis. Transcriptomics profiles of the dose-dependent response to triptolide suggested that PI3K/AKT, MAPK, TNFα and p53 signaling pathways were the vital steps in triptolide-induced hepatocyte apoptosis. Metabolomics further revealed that glycerophospholipid, fatty acid, leukotriene, purine and pyrimidine metabolism were the major metabolic alterations after triptolide exposure. Finally, acylcarnitines were identified as potential biomarkers for the early detection of triptolide-induced liver injury.

Topics: Animals; Apoptosis; Chemical and Drug Induced Liver Injury; Diterpenes; Dose-Response Relationship, Drug; Epoxy Compounds; Gene Expression Profiling; Metabolome; Metabolomics; Mice; Mice, Inbred C57BL; Mice, Knockout; Necroptosis; Phenanthrenes; Transcriptome

Triptolide (TP) is the most effective ingredient found in the traditional Chinese herbal

Topics: Animals; Apoptosis; Cell Line, Tumor; Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Furans; Glucosides; Hep G2 Cells; Humans; Liver; Mice; Mice, Inbred BALB C; NF-E2-Related Factor 2; Oxidative Stress; Phenanthrenes; Protective Agents; Signal Transduction

Tripterygium wilfordii multiglycoside( GTW),an extract derived from T. wilfordii,has been used for rheumatoid arthritis and other immune diseases in China. However its potential hepatotoxicity has not been investigated completely. Firstly,the content of triptolid( TP) in GTW was 0. 008% confirmed by a LC method. Then after oral administration of GTW( 100,150 mg·kg-1) and TP( 12 μg·kg-1) in female Wistar rats for 24 h,it was found that 150 mg·kg-1 GTW showed more serious acute liver injury than 12 μg·kg-1 TP,with the significantly increased lever of serum ALT,AST,TBA,TBi L,TG and bile duct hyperplasia even hepatocyte apoptosis. The expression of mRNA and proteins of liver bile acid transporters such as BSEP,MRP2,NTCP and OATP were down-regulated significantly by GTW to inhibit bile acid excretion and absorption,resulting in cholestatic liver injury. Moreover,GTW was considered to be involved in hepatic oxidative stress injury,although it down-regulated SOD1 and GPX-1 mRNA expression without significant difference in MDA and GSH levels. In vitro,we found that TP was the main toxic component in GTW,which could inhibit cell viability up to 80% in Hep G2 and LO2 cells at the dose of 0. 1 μmol·L-1. Next a LC-MS/MS method was used to detect the concentration of triptolid in plasma from rats,interestingly,we found that the content of TP in GTW was always higher than in the same amount of TP,suggesting the other components in GTW may affect the TP metabolism. Finally,we screened the substrate of p-glycoprotein( p-gp) in Caco-2 cells treated with components except TP extrated from GTW,finding that wilforgine,wilforine and wilfordine was the substrate of p-gp. Thus,we speculated that wilforgine,wilforine and wilfordine may competitively inhibit the excretion of TP to bile through p-gp,leading to the enhanced hepatotoxity caused by GTW than the same amount of TP.

Topics: Animals; Caco-2 Cells; Chemical and Drug Induced Liver Injury; Chromatography, Liquid; Diterpenes; Drugs, Chinese Herbal; Epoxy Compounds; Female; Glycosides; Humans; Liver; Phenanthrenes; Plant Extracts; Rats; Rats, Wistar; Tandem Mass Spectrometry; Tripterygium

To explore the protective effects of quercetin (QE) on triptolide (TP) induced liver injury and the relevant mechanism.. Forty C57BL/6 mice were equally divided into 4 groups, control group, TP model group, 20 mg/kg QE treatment group and 80 mg/kg QE treatment group randomly. The 20 mg/kg and 80 mg/kg QE groups were gastrointestinal administration with QE at the dose of 0.2 mL/10 g for 10 d, twice daily, while other groups were administrated with equivalent normal saline. Four hours post the last dose, animals were gastrointestinal administered with TP at a dose of 500 μg/kg per mouse, except for NS control. All the mice were sacrificed 22 h later, blood and liver tissue samples were collected. The pathologic change of liver tissue was detected by HE staining. The level of aminotransferase (AST) and aspartate alanine aminotransferase (ALT) in serum, and the level of glutathione (GSH), malondialdehyde (MDA) and superoxide dismutase (SOD) in liver tissue homogenates were detected using the commercial kits. The level of interleukin (IL)-17, IL-10 and IL-6 in liver tissue homogenates was measured by ELISA. Hepatic expression of Toll-like receptor 4 (TLR4) was detected by Western blot.. Compared with the control group, in the TP model group, hepatic lobule structure atrophied and even disappeared, hepatic cell necrosis and inflammatory cell infiltration are obvious. Additionally, in TP model group, serum ALT, AST and MDA levels were significantly increased, SOD and GSH levels were decreased, IL-6 and IL-17 levels were increased, IL-10 levels were decreased, and TLR4 protein levels were increased (. Quercetin can reduce TP-induced liver injury by reducing oxidative damage, promoting antioxidant and regulating cytokine secretion.

Topics: Animals; Antioxidants; Chemical and Drug Induced Liver Injury; Cytokines; Diterpenes; Epoxy Compounds; Liver; Mice; Mice, Inbred C57BL; Oxidative Stress; Phenanthrenes; Quercetin; Random Allocation

This study was designed to investigate whether the mice treated with triptolide (TP) could disrupt the liver immune homeostasis, resulting in the inability of the liver to eliminate the harmful response induced by lipopolysaccharide (LPS). In addition, we explored whether apoptosis and necroptosis played a critical role in the progression of the hepatotoxicity induced by TP-LPS co-treatment.. Female C57BL/6 mice were continuously administrated with two different doses of TP (250 μg/kg and 500 μg/kg) intragastrically for 7 days. Subsequently, a single dose of LPS (0.1 mg/kg) was injected intraperitoneally to testify whether the liver possesses the normal immune function to detoxicate the exogenous pathogen's stimulation. To prove the involvement of apoptosis and necroptosis in the liver damage induced by TP-LPS co-treatment, apoptosis inhibitor Z-VAD-FMK (FMK) and necroptosis inhibitor necrostatin (Nec-1) were applied before the stimulation of LPS to diminish the apoptosis and necroptosis respectively.. TP or LPS alone did not induce significant liver damage. However, compared with TP or LPS treated mice, TP-LPS co-treatment mice showed obvious hepatotoxicity with a remarkable elevation of serum ALT and AST accompanied by abnormal bile acid metabolism, a depletion of liver glycogen storage, aberrant glucose metabolism, an up-regulation of inflammatory cell infiltration, and an increase of apoptosis and necroptosis. Intraperitoneal injection of FMK or Nec-1 could counteract the toxic reactions induced by TP-LPS co-treatment.. TP could disrupt the immune response, resulting in hypersensitivity of the liver upon LPS stimulation, ultimately leading to abnormal liver function and cell death. Additionally, apoptosis and necroptosis played a vital role in the development of liver damage induced by TP-LPS co-treatment.

Topics: Alanine Transaminase; Amino Acid Chloromethyl Ketones; Animals; Apoptosis; Aspartate Aminotransferases; Bile Acids and Salts; Caspase Inhibitors; Chemical and Drug Induced Liver Injury; Diterpenes; Dose-Response Relationship, Drug; Epoxy Compounds; Female; Glucose; Glycogen; Imidazoles; Immunologic Factors; Indoles; Lipopolysaccharides; Liver; Mice, Inbred C57BL; Necrosis; Phenanthrenes; Signal Transduction

Triptolide (TP), as the main component of Tripterygium wilfordii (TW), could induce significant hepatic damage when exerting the therapeutic effect. However, The previous studies have shown that catalpol (CAT) and panax notoginseng saponins (PNS) have synergistic protecting effect against hepatotoxicity induced by TP. This study aims to address the role of the nuclear factor erythroid-2-related factor-2 (Nrf2)/antioxidant response element (ARE) pathway in their protecting effect, and to explore their synergistic mechanisms. For this purpose, the human hepatocyte cell line L-02 was selected, and the synergistic antioxidative effect of CAT and PNS was confirmed. Then the effects of CAT and PNS on different aspects of the Nrf2/ARE pathway were analyzed. The results showed that CAT significantly reduced TP-induced inhibition of Nrf2 transcription and made it increased, PNS significantly increased Nrf2 phosphorylation for relieving TP-induced potential inhibition of Nrf2/ARE binding activity, while the combination of the two further enhanced their activation by synergistically inducing Nrf2 transcriptional expression and phosphorylated Nrf2 (p-Nrf2) expression, followed with NAD(P)H:quinine oxidoreductase 1 (NQO1) expression and glutathione (GSH) activity increasing, which further enhanced their antioxidative effects. Subsequently, the gene silencing techniques were used, and that the Nrf2/ARE pathway is necessary in this effect was confirmed. In conclusion, the synergistic protecting effect of CAT and PNS is dependent on Nrf2/ARE pathway, while the natural mutual promotion between the regulatory links of CAT and PNS may be the main source of the synergistic effect.

Topics: Antioxidant Response Elements; Cell Line; Chemical and Drug Induced Liver Injury; Diterpenes; Drug Synergism; Epoxy Compounds; Humans; Iridoid Glucosides; NF-E2-Related Factor 2; Panax notoginseng; Phenanthrenes; Saponins; Signal Transduction

Polycyclic aromatic hydrocarbons (PAHs) are a class of pervasive global environmental pollutants and adversely affect human health. Among PAHs, phenanthrene and anthracene are isomers consisting of three benzene rings. In the present study, we have made comparisons of constitutive androstane receptor (CAR) activation and toxic effects on the liver between these two isomers. Phenanthrene, but not anthracene, significantly induced promoter activity and gene expression of human drug metabolizing enzyme CYP2B6 in HepG2 cells and human primary hepatocytes, respectively. Phenanthrene, but not anthracene, significantly increased CYP2B10 expression levels and caused hepatotoxicity in mice. Phenanthrene induced the nuclear accumulation of CAR in the liver of wild-type mice, but not CAR

Topics: Animals; Anthracenes; Aryl Hydrocarbon Hydroxylases; Cell Line, Tumor; Chemical and Drug Induced Liver Injury; Constitutive Androstane Receptor; Cytochrome P450 Family 2; Gene Expression; Glucuronosyltransferase; Glutathione Transferase; Hep G2 Cells; Hepatocytes; Humans; Liver; Male; Mice; Mice, Inbred C57BL; Phenanthrenes; Receptors, Cytoplasmic and Nuclear; Sulfotransferases

Triptolide (TP) has been widely used in China for more than 40 years as an immunosuppressive agent. Recently, serious concerns have been raised over TP-induced liver injury, though the real hepatotoxic mechanism is still unclear, particularly in terms of the initial cause. To our knowledge, this study is the first to screen systematically the mechanism of TP-induced toxicity through a global cytotoxicity profile high-content analysis using three independent cytotoxic assay panels with multiple endpoints of cytotoxicity, including cell loss, mitochondrial membrane potential, nuclear membrane permeability, manganese superoxide dismutase, phosphorylated gamma-H2AX, light chain 3B, lysosome, reactive oxygen species and glutathione. We assessed nine parameters and four stress response pathway models by labeling nuclear factor erythroid 2-related factor 2, activating transcription factor 6, hypoxia inducible factor 1α and nuclear factor κB and found that all testing parameters except glutathione and manganese superoxide dismutase showed concentration- and time-dependent changes, as well as increased cell loss after TP treatment. Considering that RNA polymerase II is the molecular target of TP, we quantified transcription from inducible genes, bromodeoxyuridine incorporation, and expression from transiently transfected green fluorescence protein plasmids in HepG2 cells. The results show that inhibition of global transcription by TP took place at earlier times and at lower concentrations than those observed for cell death. Therefore, global transcriptional suppression and the cell dysfunction it drives play a central role in TP-induced hepatotoxicity. This provides valuable information for the safe use of TP in the clinic.

Topics: Cells, Cultured; Chemical and Drug Induced Liver Injury; China; Diterpenes; Epoxy Compounds; Humans; Immunosuppressive Agents; Medicine, Chinese Traditional; Phenanthrenes; Plant Extracts; Plant Roots; Tripterygium

Triptolide, a purified diterpenoid from the herb Tripterygium wilfordii Hook.f., was widely used to treat many diseases. However, the hepatotoxicity of triptolide limited its clinical use. Research showed oxidative stress played an important role in triptolide-induced liver injury. To investigate the effect of vitamin C, which was one of the most effective antioxidants, on triptolide-induced hepatotoxicity and its potential mechanism in mice. In the present study, acute liver injury was induced by intraperitoneal injection of triptolide and vitamin C was orally administered. The results showed treatment with vitamin C prevented the triptolide-induced liver injury by reducing the levels of aspartate transaminase from 286.86 to 192.48 U/mL and alanine aminotransferase from 746.75 to 203.36 U/mL. Histopathological changes of liver corresponded to the same trend. Furthermore, vitamin C also protected the liver against triptolide-induced oxidative stress by inhibiting the generation of malondialdehyde (2.22 to 1.49 nmol/mgprot) and hydrogen peroxide (14.74 to 7.19 mmol/gprot) and restoring the level of total superoxide dismutase (24.32 to 42.55 U/mgprot) and glutathione (7.69 to 13.03 μg/mgprot). These results indicated that vitamin C could protect against triptolide-induced liver injury via reducing oxidative stress, and vitamin C may pose a significant health protection in the clinical use of triptolide.

Topics: Alanine Transaminase; Animals; Ascorbic Acid; Aspartate Aminotransferases; Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Male; Mice; Mice, Inbred C57BL; Oxidative Stress; Phenanthrenes; Protective Agents

Previous research about the development of triptolide (TP) as a natural active compound has often focused on hepatotoxicity. Among its various mechanisms, autophagy and apoptosis are two important signaling pathways. In this study, we used zebrafish to establish a TP-induced hepatotoxicity model, and investigated the roles of autophagy and apoptosis in the progress of liver injury. Zebrafish exposed to TP showed increased mortality and malformation because of the increased drug dose and duration of exposure. Meanwhile, we found that TP induced liver injury in a time- and dose-dependent manner, which was observed as a reduction in liver area, slow yolk absorption, upregulation of transaminase and local neurosis. With the application of the high-content imaging system (HCIS) technique in liver 3D imaging in vivo, clear imaging of the zebrafish liver was achieved. The results showed a decrease in volume and location of necrosis in the liver after TP exposure. Increased expression of inflammatory cytokines genes tumor necrosis factor (Tnf)α, Il1β and Il6 were shown, particularly Tnfα. The Fas-Caspase8 signaling pathway was activated. The apoptosis-related gene Bcl-2 was increased, and Bax, Caspase9 and Caspase3 were increased. However, autophagy related genes Beclin1, Atg5, Atg3 and Lc3 were increased more significantly, and the changes of Beclin1 and Atg5 were the most severe. This study successfully established a TP-induced zebrafish hepatotoxicity model and applied the HCIS technique in a zebrafish hepatotoxicity study. The result indicated Fas might be the main target of TP-induced hepatotoxicity. Autophagy played a more important role than apoptosis and was characterized by the overexpression of Beclin1 and Atg5.

Topics: Animals; Apoptosis; Autophagy; Autophagy-Related Protein 5; Beclin-1; Chemical and Drug Induced Liver Injury; Diterpenes; Dose-Response Relationship, Drug; Embryo, Nonmammalian; Embryonic Development; Epoxy Compounds; Larva; Liver; Phenanthrenes; Zebrafish; Zebrafish Proteins

Triptolide (TP), the major active compound derived from the traditional Chinese medicine Tripterygium wilfordii Hook. F, possesses an excellent pharmacological profile of immunomodulatory and anti-tumor activities. However, the application of TP was restricted due to its narrow therapeutic window and side effects, especially its hepatotoxicity. This study was designed to investigate the role of inflammasome in TP-induced acute liver toxicity. After the administration of TP at the dose of 600 μg/kg for 12 h or 24 h, we examined the serum biochemical parameters, liver histopathological changes, the expression of liver inflammatory factors, and the activation of NLRP3 inflammasome. Mice treated with TP displayed liver injury with a time-dependent increase of serum transaminases and activation of NLRP3 inflammasome, accompanied by the elevation of neutrophils infiltration. Further results implied that the activation of TLR4-Myd88-NF-κB pathway and oxidative stress induced by a single dose of TP (600 μg/kg) might participate in the activation of NLRP3 inflammasome. To investigate whether the activation of inflammasome participates in the liver damage induced by TP, a single dose of Ac-Yvad-Cmk (Caspase-1 inhibitor) was injected before TP administration. Ac-Yvad-Cmk pretreatment effectively prevented the increase of Cleaved Caspase-1 and inhibited the maturity of IL-1β. Additional studies revealed that Ac-Yvad-Cmk pretreatment decreased the recruitment of neutrophils and inhibited the production of massive pro-inflammatory factors. Taken together, our results revealed that activation of inflammasome aggravated the acute liver toxicity induced by TP. Inhibition of inflammasome could serve as a novel therapeutic target for the amelioration of TP-induced hepatotoxicity.

Topics: Acute Disease; Animals; Chemical and Drug Induced Liver Injury; Cytokines; Diterpenes; Epoxy Compounds; Female; Inflammasomes; Liver; Mice, Inbred C57BL; Myeloid Differentiation Factor 88; NF-KappaB Inhibitor alpha; NLR Family, Pyrin Domain-Containing 3 Protein; Oxidative Stress; Phenanthrenes; Toll-Like Receptor 4

Silymarin has been used in the treatment of a number of liver diseases for a long time, but its efficacy in preventing triptolide induced acute hepatotoxicity has not been reported previously. The present study aimed to assess the protective effect of silymarin against triptolide (TP)-induced hepatotoxicity in rats. Rats were orally administrated with silymarin (50, 100 and 200 mg/kg) for 7 days and received intraperitoneal TP (2 mg/kg) on the day 8. Hepatic injuries were comprehensively evaluated in terms of serum parameters, morphological changes, oxidative damage, inflammation and apoptosis. The results demonstrated that TP-induced increases in serum parameters, including alanine transaminase, aspartate aminotransferase, alkaline phosphatase, total cholesterol and γ-glutamyl transpeptidase, which were determined using a biochemical analyzer, and histopathological alterations and hepatocyte apoptosis as determined by hematoxylin and eosin and TUNEL staining, respectively, were prevented by silymarin pretreatment in a dose-dependent manner. TP-induced depletions in the activity of the antioxidant enzymes superoxide dismutase, glutathione peroxidase, glutathione S-transferase and catalase, and glutathione levels, were also significantly reversed by silymarin, as determined using specific kits. Additionally, silymarin dose-dependently exhibited inhibitory effects on malonaldehyde content in the liver. The production of proinflammatory cytokines was investigated using ELISA kits, and the results demonstrated that silymarin dose-dependently inhibited the production of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-10 and IL-1β in the liver. To determine the mechanism of silymarin, western blot analysis was performed to investigate the protein expression of phosphorylated (p)-p38 and p-c-Jun N-terminal kinase (JNK) of the TNF-α induced inflammatory response and apoptotic pathways. Silymarin significantly blocked p38 and JNK phosphorylation and activation. Additionally, the expression of the proapoptotic proteins cytochrome c, cleaved caspase-3 and Bcl-2-associated X was also reduced following treatment with silymarin, as determined by ELISA, western blotting and immunohistochemistry, respectively. In conclusion, silymarin was demonstrated to dose-dependently protect rat liver from TP-induced acute hepatotoxicity, with the high dose (200 mg/kg) achieving a superior effect. This protective effect may be associated with the improvement of antioxidant and

Topics: Animals; Antioxidants; Apoptosis; Apoptosis Regulatory Proteins; Caspase 3; Chemical and Drug Induced Liver Injury; Cytokines; Disease Models, Animal; Diterpenes; Epoxy Compounds; Hepatocytes; Inflammation Mediators; Lipid Peroxidation; Male; Oxidative Stress; Phenanthrenes; Protective Agents; Rats; Reactive Oxygen Species; Silymarin

The hepatotoxicity of Tripterygium wilfordii Hook. f. (TW), due to the presence of triptolide (TP), limits its therapeutic potential. Based on the traditional Chinese medicine theory, the theory of "Yi lei xiang zhi" was proposed that Chinese herbs with different efficacy can restrict each other to achieve the least adverse reactions.. To observe the effects of Catapol (CAT) and Panax notoginseng saponins (PNS), active ingredients in Rehmannia glutinosa (RG) and Panax notoginseng (PN) respectively, on reducing TP-induced hepatotoxicity, and further to explore the mechanisms.. The human hepatic cell line L-02 was cultured and treated with CAT, PNS or Combinations, and then treated with TP. The cytotoxic assay, the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH), apoptosis, mitochondrial membrane potential and the expressions of NF-E2-related factor 1 (Nrf1) and its downstream targets were detected. Rats were treated with TP, TP + CAT, TP + PNS, or the combinations for 4 weeks. The levels of ALT, AST and LDH in serum, apoptosis of liver cells, mitochondria injury and the protein expressions of Caspase 3 and Nrf1 were investigated.. CAT, PNS or CAT+PNS pre-treatment inhibited TP-induced toxicity in L-02 cells, distinctly decreased the apoptosis, alleviated the reduction of mitochondrial membrane potential, and modulated the expressions of Nrf1 and its downstream target, the mitochondrial transcription factor A (TFAM) and cytochrome C (Cyt-C). CAT, PNS or CAT+PNS inhibited the TP-induced hepatotoxicity in SD rats by reducing the mitochondria injury, decreasing the cells apoptosis and increasing the Nrf1 protein expression. Noticeably, TP + PNS + CAT combinations exhibited more effective than any single ingredient alone.. PNS and CAT were able to effectively attenuate TP-induced hepatotoxicity. The efficiency benefits from their modulating Nrf1 and its downstream genes TFAM and Cyt-C, and further influencing mitochondrial functions and cells apoptosis. The combination is more effective than single ingredient alone.

Topics: Animals; Apoptosis; Biomarkers; Caspase 3; Cell Line; Chemical and Drug Induced Liver Injury; Cytochromes c; Cytoprotection; Disease Models, Animal; Diterpenes; DNA-Binding Proteins; Dose-Response Relationship, Drug; Drug Therapy, Combination; Drugs, Chinese Herbal; Epoxy Compounds; Female; Humans; Liver; Membrane Potential, Mitochondrial; Mitochondria, Liver; Mitochondrial Proteins; NF-E2-Related Factor 1; Panax; Phenanthrenes; Phytotherapy; Plants, Medicinal; Quaternary Ammonium Compounds; Rats, Sprague-Dawley; Saponins; Transcription Factors

Triptolide, the predominant biologically active component of the Chinese herb Tripterygium wilfordii Hook f., possesses numerous pharmacological activities, including anti-inflammatory, anti-fertility, anti-neoplastic, and immunosuppressive effects. However, toxicity and severe adverse effects, particularly hepatotoxicity, limit the clinical application of triptolide. Licorice root extract contains various bioactive compounds and is potent hepatoprotective. Magnesium isoglycyrrhizinate, a magnesium salt of the 18α-glycyrrhizic acid stereoisomer of glycyrrhizic acid, is used clinically in China to treat chronic viral hepatitis and acute drug-induced liver injury. The aim of this study was to investigate the role of the factor erythroid 2-related factor 2 pathway in the protective effects of LE and MIG against triptolide-induced hepatotoxicity. Hepatotoxicity models were established in L-02 cells and rats using triptolide, and the protective effects of LE and MIG were investigated in vitro and in vivo, respectively. LE and MIG significantly protected against triptolide-induced cytotoxicity. Additionally, triptolide decreased the mRNA and protein levels of Nrf2 and down-regulated Nrf2 target genes, including UGT1A, BSEP, and MRP2, while pretreatment with LE and MIG reversed these effects. Finally, Nrf2-involved antioxidant responses were activated in the presence of LE and MIG.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Chemical and Drug Induced Liver Injury; Diterpenes; Down-Regulation; Drug-Related Side Effects and Adverse Reactions; Drugs, Chinese Herbal; Epoxy Compounds; Glycyrrhiza; Humans; Liver; Male; NF-E2-Related Factor 2; Oxidative Stress; Phenanthrenes; Plant Extracts; Plant Roots; Rats; Rats, Wistar; Saponins; Triterpenes; Up-Regulation

Triptolide, a major active constitute of Tripterygium wilfordii Hook. F, is prescribed for the treatment of autoimmune diseases in China. One of its most severe adverse effects observed in the clinical use is hepatotoxicity, but the mechanism is still unknown. Therefore, the present study applied an LC/MS-based metabolomic analysis to characterize the metabolomic changes in serum and liver induced by triptolide in mice. Mice were administered triptolide by gavage to establish the acute liver injury model, and serum biochemical and liver histological analyses were applied to assess the degree of toxicity. Multivariate data analyses were performed to investigate the metabolic alterations. Potential metabolites were identified using variable importance in the projection values and Student's t-test. A total of 30 metabolites were observed that were significantly changed by triptolide treatment and the abundance of 29 metabolites was correlated with the severity of toxicity. Pathway analysis indicated that the mechanism of triptolide-induced hepatotoxicity was related to alterations in multiple metabolic pathways, including glutathione metabolism, tricarboxylic acid cycle, purine metabolism, glycerophospholipid metabolism, taurine and hypotaurine metabolism, pantothenate and CoA biosynthesis, pyrimidine metabolism and amino acid metabolism. The current study provides new mechanistic insights into the metabolic alterations that lead to triptolide-induced hepatotoxicity.

Topics: Animals; Chemical and Drug Induced Liver Injury; Chromatography, High Pressure Liquid; Chromatography, Liquid; Diterpenes; Epoxy Compounds; Liver; Male; Metabolome; Metabolomics; Mice; Mice, Inbred C57BL; Phenanthrenes; Spectrometry, Mass, Electrospray Ionization

To discuss the curative effect and security of triptolide (TPL) on TNF transgenic (TNF-Tg) mice with rheumatoid arthritis (RA), and to explore the mechanism primarily.. 40 TNF-Tg RA mice were randomlydivided into five groups averagely: the control group, low-dose group (3.3 μg/kg/d TPL), middle-dose group (10 μg/kg/d TPL), high-dose group (33 μg/kg/d TPL) and MTX group (0.1 mg/kg/d MTX). Mice were administrated five days a week for six weeks. The arthritis deformation index, arthritis detumescencepercentage and the level of inflammatory factor in each group were recorded during theadministration. After administration, body weight, liver and renal function indexes, the apoptosis rates of osteoclast precursors (OCP), T and B lymphocytes in the peripheral blood and the number of osteoclast (OC) were detected and compared. μCT scanning and HE staining methods were taken to observethebone histomorphometry and bony erosion.. After administration, the arthritis deformation indexes were lower and arthritis detumescence percentageswere higher in TPL groups thanthe control group (P < 0.05), and the arthritis detumescence percentage in the high-dose group was higher than the MTX group (P < 0.05). The liver function index ALT increased after administrationin the high-dose group but was lower than that in the MTX group (P < 0.05). The level of IL-1α, IL-1β, and TNF-α decreased in the TPL groups and MTX group after administration;The apoptosis rates of OCP and T lymphocytes in middle and high dose TPL groups and MTX group were higher than other groups, and that in the high-dose group was higher than the MTX group (P < 0.05). Compared with the other groups, the bony erosion degree was lower and the number of OC was less and the parameters of bone histomorphometry were better in the high-dose group.. TPL could improvearthritic of TNF-Tg mice by decreasing the levels of pro-inflammatory cytokines, promoting the apoptosis of OCP, inhibiting the generation of OC and bone resorption. There was some toxic and side effect on liver for high-dose TPL which was weaker than the MTX.

Topics: Animals; Antirheumatic Agents; Apoptosis; Arthritis, Rheumatoid; Biomarkers; Bone Resorption; Chemical and Drug Induced Liver Injury; Cytokines; Disease Models, Animal; Diterpenes; Dose-Response Relationship, Drug; Epoxy Compounds; Inflammation Mediators; Joints; Lymphocytes; Mice, Inbred C57BL; Mice, Inbred CBA; Mice, Transgenic; Osteoclasts; Phenanthrenes; Risk Assessment; Time Factors; Tumor Necrosis Factor-alpha

Triptolide (TP) is the main active ingredient of Tripterygium wilfordii Hook.f, which has attracted great interest due to its promising efficacy for autoimmune diseases and tumors. However, severe adverse reactions, especially hepatotoxicity, have restricted its approval in the market. In the present study we explored the role of hepatic natural killer T (NKT) cells in the pathogenesis of TP-induced liver injury in mice. TP (600 μg/kg/day, i.g.) was administered to female mice for 1, 3, or 5 days. We found that administration of TP dose-dependently induced hepatotoxicity, evidenced by the body weight reduction, elevated serum ALT and AST levels, as well as significant histopathological changes in the livers. However, the mice were resistant to the development of TP-induced liver injury when their NKT cells were depleted by injection of anti-NK1.1 mAb (200 μg, i.p.) on days -2 and -1 before TP administration. We further revealed that TP administration activated NKT cells, dominantly releasing Th1 cytokine IFN-γ, recruiting neutrophils and macrophages, and leading to liver damage. After anti-NK1.1 injection, however, the mice mainly secreted Th2 cytokine IL-4 in the livers and exhibited a significantly lower percentage of hepatic infiltrating neutrophils and macrophages upon TP challenge. The activation of NKT cells was associated with the upregulation of Toll-like receptor (TLR) signaling pathway. Collectively, these results demonstrate a novel role of NKT cells contributing to the mechanisms of TP-induced liver injury. More importantly, the regulation of NKT cells may promote effective measures that control drug-induced liver injury.

Topics: Animals; Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Female; Interferon-gamma; Interleukin-4; Leukocytes; Liver; Mice, Inbred C57BL; Natural Killer T-Cells; Phenanthrenes; Signal Transduction

Acetaminophen (APAP) overdose is the most frequent cause of acute liver failure and remains a critical problem in medicine. PARP1-dependent poly(ADPribosyl)ation is a key mediator of cellular stress responses and functions in multiple physiological and pathological processes. However, whether it is involved in the process of APAP metabolism remains elusive. In this study, we find that PARP1 is activated in mouse livers after APAP overdose. Pharmacological or genetic manipulations of PARP1 are sufficient to suppress the APAP-induced hepatic toxicity and injury, as well as reduced APAP metabolism. Mechanistically, we identify pregnane X receptor (PXR) as a substrate of PARP1-mediated poly(ADP-ribosyl)ation. The poly(ADP-ribosyl)ation of PXR in ligand-binding domain activates PXR competitively and solidly, facilitates its recruitment to target gene CYP3A11 promoter, and promotes CYP3A11 gene transcription, thus resulting in increases of APAP pro-toxic metabolism. Additionally, PXR silence antagonizes the effects of PARP1 on APAP-induced hepatotoxicity. These results identifies poly(ADP-ribosyl)ation of PXR by PARP1 as a key step in APAP-induced liver injury. We propose that inhibition of PARP1-dependent poly(ADP-ribosyl)ation might represent a novel approach for the treatment of drug-induced hepatotoxicity.

Topics: Acetaminophen; Animals; Chemical and Drug Induced Liver Injury; Cytochrome P-450 CYP3A; Membrane Proteins; Mice; Mice, Inbred C57BL; Mice, Knockout; Phenanthrenes; Poly ADP Ribosylation; Poly(ADP-ribose) Polymerases; Pregnane X Receptor; Promoter Regions, Genetic; Protein Binding; RNA Interference; RNA, Small Interfering; Survival Rate; Transcriptional Activation

Hepatic macrophages are central players in the pathogenesis of some liver diseases, but few studies have examined the effect of triptolide (TP) on these cells. In this study, we investigated the possible role of hepatic macrophage recruitment and activation in triptolide-induced hepatotoxicity based on a non-hepatotoxic dose of lipopolysaccharides (LPS). The results showed that continuous administration of TP for two weeks and a single challenge of low-dose lipopolysaccharides increased the number of hepatic macrophages but inhibited their phagocytic function. TP induced the liver to recruit monocyte-derived macrophages (MoMFs) in response to a single challenge of low-dose LPS, resulting in acute inflammation and increased sensitivity to the endotoxin. Concurrent administration of TP with LPS resulted in obvious hepatotoxicity, but a single dose of LPS did not induce hepatotoxicity. These results indicate that TP could change the number and function of hepatic macrophages to reduce the ability of the liver to clear mild endotoxins, thus increasing blood endotoxin levels and increasing the sensitivity of the liver to low-dose LPS. By investigating the critical function of hepatic macrophages in TP-induced hepatotoxicity, this study elucidated the mechanism underlying TP-induced hepatotoxicity.

Topics: Animals; Cell Line; Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Lipopolysaccharides; Liver; Liver Function Tests; Macrophage Activation; Macrophages; Male; Mice, Inbred C57BL; Phagocytosis; Phenanthrenes

Triptolide (TP) induces severe liver injury, but its hepatotoxicity mechanisms are still unclear. Inflammatory responses may be involved in the pathophysiology. Neutrophils are the first-line immune effectors for sterile and non-sterile inflammatory responses. Thus, the aim of the present study was to investigate the neutrophilic inflammatory response in TP-induced liver injury in C57BL/6 mice. Our results showed that neutrophils were recruited and accumulated in the liver, which was parallel to or slightly after the development of liver injury. Neutrophils induced release of myeloperoxidase and up-regulation of CD11b, which caused cytotoxicity and hepatocyte death. Hepatic expressions of CXL1, TNF-α, IL-6, and MCP1 were increased significantly to regulate neutrophils recruitment and activation. Up-regulation of toll like receptors 4 and 9 also facilitated neutrophils infiltration. Moreover, neutrophils depletion using an anti-Gr1 antibody showed mild protection against TP overdose. These results indicated that neutrophils accumulation might be the secondary response, not the cause of TP-induced liver injury. In conclusion, the inflammatory response including neutrophil infiltration may play a role in TP-induced hepatotoxicity, but may not be severe enough to cause additional liver injury.

Topics: Animals; Chemical and Drug Induced Liver Injury; Chemokine CCL2; Diterpenes; Drugs, Chinese Herbal; Epoxy Compounds; Female; Humans; Interleukin-6; Intracellular Signaling Peptides and Proteins; Liver; Mice; Mice, Inbred C57BL; Neutrophil Infiltration; Neutrophils; Phenanthrenes; Tripterygium; Tumor Necrosis Factor-alpha

Triptolide (TP) has outstanding biological activities, but it induces toxicities, particular hepatotoxicity, severely limiting its clinical application. Chlorogenic acid (CGA) has prominently medicinal and nutritional values. However, until now, it is not known whether CGA could mitigate TP-induced hepatotoxicity.. This study explored the possible protection of CGA against TP-induced hepatotoxicity and its potential mechanisms, for the first time.. KM mice were treated orally with TP at a single dose of 1 mg/kg at 4 h after being treated with CGA (10, 20 and 40 mg/kg) for seven continuous days. Blood samples were collected at 24 h after TP administration for measurement of serum biomarkers, and hepatic tissues for analysis of potential mechanisms.. Combined CGA medication may probably reduce the risk of TP poisoning, and in-depth mechanisms can be developed around the signal molecules of Nrf2.

Topics: Animals; Antineoplastic Agents, Alkylating; Chemical and Drug Induced Liver Injury; Chlorogenic Acid; Diterpenes; Dose-Response Relationship, Drug; Epoxy Compounds; Male; Mice; Oxidative Stress; Phenanthrenes; Protective Agents

Triptolide is a vine extract used in traditional Chinese medicines and associated with hepatotoxicity. In vitro data suggest that inhibition of RNA synthesis may be the mechanism of toxicity. For studying drug-induced liver injury the zebrafish has experimental, practical and financial advantages compared with rodents. The aim of this study was to explore the mechanism of triptolide toxicity using zebrafish as the model system. The effect of triptolide exposure on zebrafish larvae was determined with regard to mortality, histology, expression of liver specific microRNA-122 and liver volume. Fluorescent microscopy was used to track toxicity in the Tg(-2.8lfabp:GFP)as3 zebrafish line. Informed by microscopy, RNA-sequencing was used to explore the mechanism of toxicity. Triptolide exposure resulted in dose-dependent mortality, a reduction in the number of copies of microRNA-122 per larva, hepatocyte vacuolation, disarray and oncotic necrosis, and a reduction in liver volume. These findings were consistent across replicate experiments. Time-lapse imaging indicated the onset of injury was 6 h after the start of exposure, at which point, RNA-sequencing revealed that 88% of genes were down-regulated. Immune response associated genes were up-regulated in the triptolide-treated larvae including nitric oxide synthase. Inhibition of nitric oxide synthase increased mortality. Triptolide induces hepatotoxicity in zebrafish larvae. This represents a new model of drug-induced liver injury that complements rodents. RNA sequencing, guided by time-lapse microscopy, revealed early down-regulation of genes consistent with previous invitro studies, and facilitated the discovery of mechanistic inflammatory pathways.

Topics: Animals; Chemical and Drug Induced Liver Injury; Disease Models, Animal; Diterpenes; Epoxy Compounds; Larva; Liver; MicroRNAs; Microscopy, Fluorescence; Phenanthrenes; Polymerase Chain Reaction; Sequence Analysis, RNA; Transcriptome; Zebrafish

Triptolide (TP) is a diterpene triepoxide with various biological activities, but its clinical applications have been limited by potential hepatotoxicity, which can be attributed to T helper 17 (Th17)/T regulatory (Treg) cell imbalance. Quercetin (QE), a natural flavonoid, has been reported to have many benefits and medicinal properties, including hepatoprotective activity against TP-induced liver injury. However, the hepatoprotection mechanisms have not been clarified. The present study was designed to explore the protective effect and the mechanism of QE against TP-induced liver injury. Treatment with QE (20, 50 and 80mg/kg) prior to TP administration restored TP-induced alterations in a certain dose range indicating that QE was able to inhibit TP-induced liver injury. One mechanism underlying this effect was the shifting balance in Th17 and Treg cells from Th17 dominance to Treg dominance. Furthermore, QE markedly decreased the expression level of the Th17-related pro-inflammatory cytokines interleukin (IL)-17 and IL-6, as well as the Th17 transcription factor retinoid-related orphan receptor-γt (ROR-γt). TP induced downregulation in the expression of anti-inflammatory cytokine IL-10, but the expression of Treg transcription factor forkhead/winged-helix family transcriptional repressor p3 (FoxP3) was restored by QE. In the process of exploring the possible hepatoprotective mechanisms of QE, we found that QE significantly reduced both protein and mRNA expression of Toll-like receptor 4 (TLR4), which in turn not only inactivated myeloid differentiation primary response gene 88 (MYD88), nuclear factor kappa B (NF-κB) and related inflammatory cytokines IL-6 and IL-17, but also simultaneously increased the levels of T-cell immunoglobulin and mucin domain-containing protein 3 (Tim-3). Furthermore, blocking of TLR4 enhanced the effect of QE in regulating the Th17/Treg imbalance. In summary, this report has demonstrated for the first time that the protection afforded by QE against TP-induced liver injury was associated with a shift in the balance of Th17 and Treg cells to Treg dominance, which was regulated by Tim-3 and TLR4-MyD88-NF-κB signaling pathway.

Topics: Animals; Antioxidants; Cells, Cultured; Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Female; Forkhead Transcription Factors; Hepatitis A Virus Cellular Receptor 2; Liver; Mice; Mice, Inbred C57BL; Myeloid Differentiation Factor 88; NF-kappa B; Phenanthrenes; Quercetin; Signal Transduction; T-Lymphocytes, Regulatory; Th17 Cells; Toll-Like Receptor 4

Isoliquiritigenin, a flavonoid found in licorice, has been considered as an antioxidive and hepato-protective agent. Recent studies have shown that a possible mechanism for triptolide-induced hepatotoxicity is related to oxidative damage induced by reactive oxygen species. This study was done to investigate the protection effect of isoliquiritigenin against triptolide-induced hepatotoxicity and the mechanism involved. An acute liver injury model was established by intraperitoneal injection of triptolide (1.0 mg · kg-1) in mice. Different doses of isoliquiritigenin (12.5, 25 and 50 mg · kg-1) were employed as protection. The activities of AST, ALT, ALP and LDH in serum and levels of GSH, GPx, SOD, CAT and MDA in liver tissue were detected. The histopathological changes of liver tissues were observed after HE staining. The protein expression of Nrf2 was detected by western blot. Pretreatment with isoliquiritigenin significantly prevented the triptolide-induced hepatotoxicity indicated by reduced activities of AST, ALT, ALP and LDH. Moreover, isoliquiritigenin pretreatment also prevented from triptolide-induced hepatotoxicity by inhibiting MDA and restoring the levels of GSH, GPx, SOD and CAT. In addition, isoliquiritigenin could attenuate histopathological changes induced by triptolide. Furthermore, the results indicated that isoliquiritigenin pretreatment caused an increase in the protein expression of Nrf2. These results indicated that isoliquiritigenin could protect against triptolide-induced hepatotoxicity via activation of the Nrf2 pathway.

Topics: Animals; Chalcones; Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Liver; Liver Function Tests; Male; Malondialdehyde; Mice; Mice, Inbred ICR; NF-E2-Related Factor 2; Oxidative Stress; Phenanthrenes; Protective Agents; Signal Transduction

Tripterygium wilfordii Hook F (TWHF) is a traditional herbal medicine in China. Triptolide (TP), the primary bioactive compound of TWHF, is an anti-inflammatory and immunosuppressive compound that can also injure the liver and kidney. Unfortunately, the toxicity mechanism remains unknown.. The aim of this study is to understand the regulatory role of sphingolipid (SPL) pathways in the TP-induced toxic mechanism in the liver and kidney in delayed-type hypersensitivity (DTH) Balb\\c mouse.. 76 core sphingolipids and 29 species of related metabolic enzymes in liver, kidney and plasma were analyzed with previous HPLC-MS/MS and real time qPCR method, respectively. Furthermore, the data generated from these two omics underwent integrated analysis to describe TP-induced abnormal sphingolipid metabolism and identify the specific biomarkers of TP toxicity using bioinformation method.. High-dose (LD50) TP could induce severe liver and kidney injuries. Moreover, TP comprehensively influenced the enzymes involved in the sphingolipids metabolism in the liver and kidney at the mRNA expression level. Furthermore, the total levels of ceramides (Cers), sphingomyelins (SMs) and sphingosine (Sph) were all elevated, while dihydroceramides (dhCers) and hexosylceramides (HexCers) were all down-regulated. Several enzymes, including kdsr, CerS2, CerS4, CerS5 and CerS6 in the liver and Cerk in the kidney were probably responsible for the TP-induced toxic effect, identifying them as possible novel therapeutic targets. Besides, fractions of long chain SPL (C16-C20) exhibited significant increase, and fractions of unsaturated dhCer and Cer were significantly changed, both of which above may be due to the change of mRNA expression level of CerSs. Moreover, several biomarkers for the diagnosis of TP poisoning were discovered.. In summary, the regulation of SPL metabolism uncovered a novel mechanism underlying TP poisoning in the liver and kidney. In addition, key biomarkers and enzymes may play an important role in reducing the clinical risk associated with the use of TP.

Topics: Animals; Biomarkers; Chemical and Drug Induced Liver Injury; Chromatography, High Pressure Liquid; Diterpenes; Epoxy Compounds; Ethnopharmacology; Kidney Diseases; Lethal Dose 50; Medicine, Chinese Traditional; Mice; Mice, Inbred BALB C; Phenanthrenes; Real-Time Polymerase Chain Reaction; Sphingolipids; Tandem Mass Spectrometry; Transcriptome; Tripterygium

Triptolide (TP) is the major active principle of Tripterygium wilfordii Hook f. and very effective in treatment of autoimmune diseases. However, TP induced hepatotoxicity limited its clinical applications. Our previous study found that TP was a substrate of P-glycoprotein and its hepatobiliary clearance was markedly affected by P-gp modulation in sandwich-cultured rat hepatocytes. In this study, small interfering RNA (siRNA) and specific inhibitor tariquidar were used to investigate the impact of P-gp down regulation on TP-induced hepatotoxicity. The results showed that when the function of P-gp was inhibited by mdr1a-1 siRNA or tariquidar, the systemic and hepatic exposures of TP were significantly increased. The aggravated hepatotoxicity was evidenced with the remarkably lifted levels of serum biomarkers (ALT and AST) and pathological changes in liver. The other toxicological indicators (MDA, SOD and Bcl-2/Bax) were also significantly changed by P-gp inhibition. The data analysis showed that the increase of TP exposure in mice was quantitatively correlated to the enhanced hepatotoxicity, and the hepatic exposure was more relevant to the toxicity. P-gp mediated clearance played a significant role in TP detoxification. The risk of herb-drug interaction likely occurs when TP is concomitant with P-gp inhibitors or substrates in clinic.

Topics: Alanine Transaminase; Animals; Apoptosis Regulatory Proteins; Aspartate Aminotransferases; ATP Binding Cassette Transporter, Subfamily B; Cell Line, Tumor; Chemical and Drug Induced Liver Injury; Digoxin; Diterpenes; Epoxy Compounds; Gene Knockdown Techniques; Immunologic Factors; Male; Mice, Inbred BALB C; Molecular Docking Simulation; Oxidative Stress; Phenanthrenes; Protein Binding; Quinolines; RNA, Small Interfering

Triptolide, an active ingredient extracted from the Chinese herb Tripterygium wilfordii Hook f., has multiple pharmacological properties, including anti-inflammatory, immune-modulatory, and anti-proliferative activities. However, the hepatotoxicity of triptolide always limits its clinical applications.. Farnesoid X receptor (FXR) is a ligand-activated transcription factor that plays a key role in hepatoprotection through the maintenance of liver metabolism homeostasis. This study explored the role of FXR in triptolide-induced cytotoxicity and investigated whether activation of FXR can protect against triptolide-induced liver injury.. The role of FXR in triptolide-induced cytotoxicity was investigated in HepG2 cells. In addition, the protective effect of the selective FXR agonist GW4064 on triptolide-induced hepatotoxicity was explored in BALB/c mice.. HepG2 cells were transient transfected with FXR expression plasmid or FXR-siRNA. The cytotoxicity was compared using the MTT assay. The extent of liver injury was assessed by histopathology and serum aminotransferases. The expression of FXR and its target genes were detected by Western blot and qRT-PCR.. The transient overexpression of FXR protected against triptolide-induced cell death, whereas FXR knockdown with a specific small interfering RNA resulted in increased cytotoxicity. In BALB/c mice, treatment with the FXR agonist GW4064 attenuated triptolide-induced liver dysfunction, structural damage, glutathione depletion and lipid peroxidation. Moreover, the livers of GW4064-treated mice showed increased expression of FXR and several related target genes involved in phase II and phase III xenobiotic metabolism.. Taken together, these results indicate that activation of FXR attenuates triptolide-induced hepatotoxicity and provide direct implications for the development of novel therapeutic strategies against triptolide-induced hepatotoxicity.

Topics: Animals; Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Gene Knockdown Techniques; Hep G2 Cells; Humans; Isoxazoles; Liver; Male; Mice, Inbred BALB C; Phenanthrenes; Receptors, Cytoplasmic and Nuclear; RNA, Small Interfering

Drug-induced hepatotoxicity is a worldwide health issue. And diagnosing the injury in the early stage is still a challenge in clinic. In this study, pattern recognition analysis of the ultra high performance liquid chromatography-mass spectrometry (UPLC-MS) of hepatocytes HL7702 was performed to develop differential metabolites related to hepatotoxicity induced by hepatotoxicants, including carbon tetrachloride (CCl4), acetaminophen (APAP), emodin, aristolochic acid (AA) and triptolide. Hepatocytes injuries were induced by 48 h of treatment with CCl4 (4 mmol/L), APAP (6.5 mmol/L), emodin (14 μmol/L), AA (35 μmol/L) and triptolide (18 nmol/L), separately. Global metabolomics profiling, multivariate analysis and database searching were performed to discover common differential metabolites for live injury. The positive hepatoprotective drug, bifendate, was used to repair triptolide induced hepatocytes injury, and bifendate-induced changes of hepatotoxicity-related metabolites were investigated. In the results, fatty acid oxidation and cellular oxidative stress-related metabolites, including nicotinamide adenine dinucleotide and glutathione were significantly changed between the control and hepatotoxicant-treated groups, and after treatment with bifendate, those perturbed metabolites all partly returned to normal level. In conclusion, we discovered potential hepatotoxicity-related metabolites that could be used to evaluate hepatotoxicity induced by chemicals, drugs and traditional Chinese medicines. This study also proved that metabolomics is one of the effective tools to investigate drug-induced hepatotoxicity.

Topics: Acetaminophen; Aristolochic Acids; Carbon Tetrachloride; Cells, Cultured; Chemical and Drug Induced Liver Injury; Chromatography, High Pressure Liquid; Diterpenes; Emodin; Epoxy Compounds; Hepatocytes; Humans; Mass Spectrometry; Metabolomics; Phenanthrenes

Cryptotanshinone from Salvia miltiorrhiza Bunge was investigated for hepatoprotective effects in d-galactosamine (GalN)/lipopolysaccharide (LPS)-induced fulminant hepatic failure. Cryptotanshinone (20 or 40 mg/kg) was orally administered 12 and 1h prior to GalN (700 mg/kg)/LPS (10 μg/kg) injection. The increased mortality and TNF-α levels by GalN/LPS were declined by cryptotanshinone pretreatment. In addition, cryptotanshinone attenuated GalN/LPS-induced apoptosis, characterized by the blockade of caspase-3, -8, and -9 activation, as well as the release of cytochrome c from the mitochondria. In addition, cryptotanshinone significantly suppressed JNK, ERK and p38 phosphorylation induced by GalN/LPS, and phosphorylation of TAK1 as well. Furthermore, cryptotanshinone significantly inhibited the activation of NF-κB and suppressed the production of proinflammatory cytokines. These findings suggested that hepatoprotective effect of cryptotanshinone is likely associated with its anti-apoptotic activity and the down-regulation of MAPKs and NF-κB associated at least in part with suppressing TAK1 phosphorylation.

Topics: Animals; Anti-Inflammatory Agents; Apoptosis; Aspartate Aminotransferases; Caspases; Chemical and Drug Induced Liver Injury; Cytochromes c; Galactosamine; Lipopolysaccharides; Liver; Liver Failure, Acute; Male; MAP Kinase Kinase Kinases; Mice; Mice, Inbred C57BL; Mitochondria; Mitogen-Activated Protein Kinases; NF-kappa B; Phenanthrenes; Phytotherapy; Plant Extracts; Salvia miltiorrhiza; Tumor Necrosis Factor-alpha

The study was designed to investigate the immune-modulatory effects of triptolide (TP) on CD4(+) T cell responses during liver injury. Previous studies have suggested that TP plays a critical role in modulating both innate and adaptive immune reactions, but its effects on the Th17/Treg balance during TP-induced liver injury remain unknown. In this study, female C57BL/6 mice were administered by oral gavage with TP at a dose of 250 or 500 μg/kg per mouse. We examined the plasma biochemical parameters, histopathological changes, hepatic frequencies of Th17 cells and Treg cells, hepatic expression of transcriptional factors and cytokine genes and hepatic interleukin (IL)-17 and IL-10 levels in TP-administered mice. Mice treated with TP displayed liver injury with markedly increased plasma transaminase as well as hepatic mRNA expression of retinoid related orphan receptor (ROR)-γt and hepatic IL-17 level at 24h. However, hepatic frequencies of Tregs and hepatic expression of forkhead/winged-helix family transcriptional repressor p3 (FoxP3) decreased at 24h after TP administration. Furthermore, we found that elevated serum biochemical parameters positively correlated with the Th17/Treg ratios. Taken together, these results revealed a novel and interesting phenomenon of TP in the enhancement of the expansion of Th17 cells and suppression of the production of Tregs during liver injury, which may represent a new pathogenesis of TP-induced liver injury.

Topics: Animals; Chemical and Drug Induced Liver Injury; Diterpenes; Dose-Response Relationship, Drug; Epoxy Compounds; Female; Immunosuppressive Agents; Interleukin-10; Interleukin-17; Liver; Mice; Mice, Inbred C57BL; Phenanthrenes; T-Lymphocytes, Regulatory; Th17 Cells; Transcription Factors

Triptolide, the major active component of Tripterygium wilfordii Hook f. (TWHF), has a wide range of pharmacological activities. However, the toxicities of triptolide, particularly the hepatotoxicity, limit its clinical application. The hepatotoxicity of triptolide has not been well characterized yet. The aim of this study was to investigate the role of NF-E2-related factor 2 (Nrf2) in triptolide-induced toxicity and whether activation of Nrf2 could protect against triptolide-induced hepatotoxicity. The results showed that triptolide caused oxidative stress and cell damage in HepG2 cells, and these toxic effects could be aggravated by Nrf2 knockdown or be counteracted by overexpression of Nrf2. Treatment with a typical Nrf2 agonist, sulforaphane (SFN), attenuated triptolide-induced liver dysfunction, structural damage, glutathione depletion and decrease in antioxidant enzymes in BALB/C mice. Moreover, the hepatoprotective effect of SFN on triptolide-induced liver injury was associated with the activation of Nrf2 and its downstream targets. Collectively, these results indicate that Nrf2 activation protects against triptolide-induced hepatotoxicity.

Topics: Animals; Anticarcinogenic Agents; Antineoplastic Agents, Alkylating; Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Hep G2 Cells; Humans; Isothiocyanates; Mice; Mice, Inbred BALB C; NF-E2-Related Factor 2; Oxidative Stress; Phenanthrenes; Sulfoxides

The aim of this study is to investigate the protection effect of tanshinone IIA (Tan) against triptolide (TP)-induced liver injury and the mechanisms involved. Acute liver injury was induced by intraperitoneal injection of TP (1 mg x kg(-1)) in mice. The activities of AST, ALT and LDH in serum and the levels of GSH, GST, GSH-PX, SOD, CAT and MDA in liver tissue were detected. The histopathological changes of liver tissues were observed after HE staining. Nrf2 translocation in liver tissue was detected by Western blotting, and real-time PCR was used to measure the expression levels of GCLC, NQO1 and HO-1 mRNA. The results showed that pretreatment with Tan significantly prevented the TP induced liver injury as indicated by reducing the activities of AST, ALT and LDH (P < 0.01). Tan pretreatment also prevented TP-induced oxidative stress in the mice liver by inhibiting MDA and restoring the levels of GSH, GST, SOD and CAT (P < 0.05). Parallel to these changes, pretreatment with Tan could attenuate histopathologic changes induced by TP. Furthermore, the results indicated that Tan pretreatment caused nuclear accumulation of Nrf2 as well as induction of mRNA expression of antioxidant response element (ARE)-driven genes such as GCLC, NQO1 and HO-1. These results indicated that Tan could protect against TP-induced acute liver injury via the activation of Nrf2/ARE pathway.

Topics: Abietanes; Animals; Antioxidant Response Elements; Chemical and Drug Induced Liver Injury; Diterpenes; Drugs, Chinese Herbal; Epoxy Compounds; Glutamate-Cysteine Ligase; Heme Oxygenase-1; Liver; Male; Membrane Proteins; Mice; Mice, Inbred C57BL; NAD(P)H Dehydrogenase (Quinone); NF-E2-Related Factor 2; Phenanthrenes; RNA, Messenger; Signal Transduction

Triptolide, a diterpene triepoxide, is one of the major components of most functional extracts of Tripterygium wilfordii Hook f, which is known to have various biological effects, including immunosuppressive, anti-inflammatory and anti-tumor functions. We studied the inhibitory effect of triptolide on endotoxemia (ETM)-induced oxidative stress, which was induced in C57BL/6 mice by lipopolysaccharide (LPS) and D-galactosamine (D-GalN). Pretreatment with triptolide decreased the reactive oxygen species (ROS) levels, mortality rate and liver injury after LPS/D-GalN injection. We utilized comprehensive proteomics to identify alterations in liver protein expression during pretreatment with triptolide or N-acetylcysteine (NAC) after LPS/D-GalN injection, 44 proteins were found to be related to oxidative stress, mitochondria, metabolism and signal transduction, and 23 proteins of them seemed to be significantly up- or down-regulated. Furthermore, both triptolide and NAC inhibited activation of c-jun NH2-terminal kinases (JNK) and mitogen-activated protein kinase p38 (p38), phosphorylation of inhibitor of nuclear factor-kappa B (IκB) and activation of nuclear factor-κB (NF-κB). These results demonstrated that triptolide inhibited the activation of JNK and p38 by decreasing ROS levels, which in turn inhibited the hepatic injury. In addition, we set and validated the phosphorylation model of extracellular signal-regulated kinase (ERK) and proposed that triptolide probably induced ERK phosphorylation through inhibiting its dephosphorylation rates. These results showed that triptolide can effectively reduce the oxidative stress and partially rescue the damage in the liver induced by LPS/D-GalN.

Topics: Animals; Antioxidants; Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Galactosamine; Lipopolysaccharides; Liver; Male; Mice; Mice, Inbred C57BL; Mitogen-Activated Protein Kinases; NF-kappa B; Oxidative Stress; Phenanthrenes; Proteomics; Reactive Oxygen Species

Triptolide, a diterpenoid epoxide, is one of the major active ingredients of Tripterygium wilfordii Hook F, a woody vine plant called lei gong teng in China, which is used in traditional Chinese Medicine (TCM) for treating many diseases. In this paper, we investigate the relation between inhibition of mitochondrial respiratory chain and liver injury induced by triptolide. Results indicate that the secondary β-oxidation impairment caused by inhibition of mitochondrial respiratory chain is involved in triptolide-induced liver injury, which featured by microvesicular steatosis, hyperlactacidemia and enhanced oxidant stress, although other mechanisms of triptolide-induced liver injury may also exist.

Topics: Animals; Cell Respiration; Chemical and Drug Induced Liver Injury; Diterpenes; Epoxy Compounds; Fatty Liver; Female; Lactic Acid; Liver; Mitochondrial Membranes; Oxidation-Reduction; Oxidative Stress; Phenanthrenes; Plant Extracts; Rats; Rats, Sprague-Dawley; Tripterygium

Salvia miltiorrhiza is traditionally used to treat liver disease in Asia. In this study, we tested the ability of a purified extract of S. miltiorrhiza (PF2401-SF) and its constituents, tanshinone I, tanshinone IIA, and cryptotanshinone, to protect against acute and subacute liver damage induced by carbon tetrachloride by measuring serum transaminase levels, the reduced form of glutathione (GSH), antioxidant enzyme activities, and lipid peroxidation levels in the liver. We also evaluated their ability to protect primary cultured rat hepatocytes from tertiary-butylhydroperoxide (tBH) or d-galactosamine (GalN). PF2401-SF was protective at 50-200mg/kg per day in acute liver injury and 25-100mg/kg per day in subacute liver injury. Tanshinone I, tanshinone IIA, and cryptotanshinon (40 microM), inhibited lactate dehydrogenase leakage, GSH depletion, lipid peroxidation and free radical generation in vitro. PF2401-SF and its major constituents, tanshinone I, tanshinone IIA and cryptotanshinone, can protect against liver toxicity in vivo and in vitro due to its antioxidant effects.

Topics: Abietanes; Animals; Antioxidants; Carbon Tetrachloride; Chemical and Drug Induced Liver Injury; Dose-Response Relationship, Drug; Hepatocytes; Male; Phenanthrenes; Plant Extracts; Rats; Rats, Sprague-Dawley; Salvia miltiorrhiza

To analyze the gene expression profiles of mice livers injured by Leigongteng and explore the relationship between the differentially expressed genes and liver damage.. The experimental mice were randomly divided into a control group and a liver-injured group in which the mice were administrated 33 mu gamma of triptolide/kg per day for 30 d. Liver mRNAs were extracted from animals in both groups and were reverse-transcribed to cDNA with dUTP labeled by different fluorescence (Cy3, Cy5) as hybridization probes. The mixed probes were hybridized with oligonucleotide microarray chips. The fluorescent signal results were acquired by scanner and analyzed with software.. Among the 35852 target genes, 29 genes were found to be significantly differentially expressed, with 20 genes up-regulated and 9 genes down-regulated. The reliability of the differentially expressed genes was validated by RT-PCR experiments of 5 randomly selected differentially expressed genes.. Based on the biological functions of the differentially expressed genes, it is obvious that the occurrence and development of liver damage induced by Leigongteng in mice are highly associated with immune response, metabolism, apoptosis and the cell skeleton of liver cells. This might be important for elucidating the regulatory network of gene expression associated with liver damage and it may also be important for discovering the pathogenic mechanisms of liver damage induced by Leigongteng.

Topics: Animals; Chemical and Drug Induced Liver Injury; Diterpenes; Drugs, Chinese Herbal; Epoxy Compounds; Gene Expression Profiling; Gene Expression Regulation; Male; Mice; Oligonucleotide Array Sequence Analysis; Phenanthrenes; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tripterygium

Although the benefit/risk ratio of older antimalarial drugs (quinine, chloroquine) is well established less is known concerning the data about newer drugs. This article assesses and analyses the antimalarial-induced ADRs reported to the French pharmacovigilance system.. All cases of ADR(s) reported to the French pharmacovigilance database over a period of 5 years, from January 1, 1996 to December 31, 2000. Our study included the antimalarials (excluding doxycycline) used in France for the cure and prophylaxis of malaria. For each notification we noted the data relative to the patient (gender, age), the antimalarial drug (prevention or cure), the associated drugs and the adverse events (imputability, delay before onset, severity and evolution).. Between 1996 and 2000, a total of 508 reports were collected, representing 0.6% of the notifications reported to the French pharmacovigilance database over the same period. For chemoprophylaxis, the chloroquine-proguanil combination predominated (n=230, i.e. 54%), followed by mefloquine (n=163, i.e. 38%) and chloroquine (n=36, i.e. 8%). Women predominated (53%). The mean age of the patients was of 40.6+/-16.1 years (range: 0-77 years). The majority of cases (67%) were aged 26 to 60 years (n=337). For curative treatment, halofantrine was the first line drug (n=30, i.e. 38%), followed by mefloquine (n=20, i.e. 26%), quinine (n=18, i.e. 24%) and chloroquine (n=9, i.e. 12%). Whatever the indication, the chloroquine-proguanil combination (n=230) and mefloquine (n=183) represented 81% of the reports (45 and 36% respectively). We noted 1 040 adverse events corresponding to 508 observations. The adverse events were classified as severe in 41.4% of cases. However, the percentage was highest for the curative (64%) than for the prophylactic treatments (37.5%) (p<0.01). For halofantrine and quinine, the adverse events were classified as severe in respectively 76 and 67% of cases. We studied the profile of adverse events of each antimalarial drug.. Our study underlined several elements: the considerable number of psychiatric problems related to the use of chloroquine-proguanil and the hepatic disorders due to halofantrine, the profile of the adverse events of each drug and the unexpected adverse events which should not be neglected in some cases.

Topics: Adolescent; Adult; Adverse Drug Reaction Reporting Systems; Age Factors; Aged; Antimalarials; Chemical and Drug Induced Liver Injury; Child; Child, Preschool; Chloroquine; Female; France; Humans; Infant; Infant, Newborn; Male; Middle Aged; Phenanthrenes; Proguanil; Psychoses, Substance-Induced; Sex Factors

We recently reported that (Lamiaceae) may alleviate CCl(4)-induced acute hepatotoxicity in rats, possibly blocking the formation of free radicals generated during CCl(4) metabolism. Carnosol, one of the main constituents of Rosmarinus, has been shown to have antioxidant and scavenging activities. Therefore, it is plausible to expect that carnosol may mediate some of the effects of Rosmarinus on oxidative stress consequences induced by CCl(4) in the liver.. We evaluated the effectiveness of carnosol to normalize biochemical and histological parameters of CCl(4)-induced acute liver injury.. Male Sprague Dawley rats (n = 5) injured by CCl(4) (oral dose 4 g/kg of body weight) were treated with a single intraperitoneal dose (5 mg/kg) of carnosol. Twenty-four hours later, the rats were anaesthetized deeply to obtain the liver and blood, and biochemical and histological parameters of liver injury were evaluated.. Carnosol normalized bilirubin plasma levels, reduced malondialdehyde (MDA) content in the liver by 69%, reduced alanine aminotransferase (ALT) activity in plasma by 50%, and partially prevented the fall of liver glycogen content and distortion of the liver parenchyma.. Carnosol prevents acute liver damage, possibly by improving the structural integrity of the hepatocytes. To achieve this, carnosol could scavenge free radicals induced by CCl(4), consequently avoiding the propagation of lipid peroxides. It is suggested that at least some of the beneficial properties of Rosmarinus officinalis are due to carnosol.

Topics: Abietanes; Acute Disease; Animals; Carbon Tetrachloride; Chemical and Drug Induced Liver Injury; Disease Models, Animal; Hepatocytes; Liver; Liver Diseases; Male; Phenanthrenes; Rats; Rats, Sprague-Dawley

To study the effect of tanshinone IIA on two models of acute hepatic injury in mice.. Two acute hepatic injury models induced by carbon tetrachloride (CCl4) and D-galactosamine (D-GalN) in mice were used. ALT, AST in serum and MDA in liver homogenate were used as indexes.. Tanshinone IIA (10 mg.kg-1, 20 mg.kg-1, 30 mg.kg-1), similar to biophenyldicarboxylate, could decrease ALT, AST in serum and MDA in hepatic homogenate on acute hepatic injury induced by CCl4 and D-GalN.. Tanshinone IIA possessed obvious protective effect against liver injury induced by CCl4 or D-GalN.

Topics: Abietanes; Animals; Carbon Tetrachloride Poisoning; Chemical and Drug Induced Liver Injury; Male; Mice; Phenanthrenes; Phytotherapy

We encountered two cases of Chinese herb-induced Fanconi syndrome in Japan. One component of the chinese medicine was "Kan-mokutsu" (Aristolochia manshuriensis) in which aristolochic acids (AAs) were detected.. Renal biopsy showed flattening of proximal tubular epithelial cells and paucicellular interstitial fibrosis without glomerular lesions, all of which were in accordance with Chinese herb nephropathy (CHN). To date, many cases of CHN have been reported mainly as progressive renal failure in western countries.. However, our cases were different from those in that they presented Fanconi syndrome. The detected AAs in our cases consisted of aristolochic acid (AA)-I, II and D. In contrast, in Belgium, the incriminated agent was Aristolochia fangchi which consisted of AA-I, B, C, and aristolactum.. These findings could indicate that different components of AAs could cause different clinical lesions, or that the amount of ingested AAs might reflect clinical pictures, that is to say, our patients took lower volume of Chinese herbs and might be in an early stage of CHN. Furthermore, it is likely that susceptibility to this substance may be different among races. CHN would include two clinical aspects: subacute renal failure and adult-onset Fanconi syndrome. It is important to bear in mind that CHN could present Fanconi syndrome.

Topics: Adult; Aristolochic Acids; Chemical and Drug Induced Liver Injury; Drugs, Chinese Herbal; Fanconi Syndrome; Female; Humans; Japan; Male; Middle Aged; Phenanthrenes

Male Sprague-Dawley rats were treated ip with beta-naphthoflavone (BNF, 40 mg/kg/day) in dimethylsulfoxide (DMSO, 26.7 mg BNF/ml) for three days. At 24 hr after the pretreatment DMSO (3.0 ml/kg), phenanthrene (150 mg/kg), ozonized or nitrated products of phenanthrene (150 mg/kg), pyrene (150 mg/kg), or ozonized or nitrated products of pyrene (150 mg/kg) were injected ip. Phenanthrene, pyrene, and their ozonized or nitrated products were dissolved in DMSO (50 mg/ml). No increase in the level of aspartate aminotransferase (AST), alanine aminotransferase (ALT) or sorbitol dehydrogenase (SDH) was seen in the pretreated rats 48 hr after the treatment. This is in contrast to what was seen in previous work without the BNF pretreatment. BNF pretreatment induced a small but significant increase in gamma-glutamyl transpeptidase (GGTP) levels. No treatment group receiving BNF differed from another with respect to GGTP. A decrease in lactate dehydrogenase (LDH) levels was noted in the nitro-PAH treatment groups; the same phenomenon was observed earlier in rats treated with nitro-PAH without BNF treatment. These results suggest that the mixed-function oxidase systems specifically induced by BNF have a protective effect against the hepatotoxicity of the oxonized or nitrated products of phenanthrene and pyrene.

Topics: Animals; Benzoflavones; beta-Naphthoflavone; Chemical and Drug Induced Liver Injury; Flavonoids; L-Lactate Dehydrogenase; Liver Diseases; Male; Nitrogen Dioxide; Ozone; Phenanthrenes; Pyrenes; Rats; Rats, Inbred Strains

An h.p.l.c.-fluorescence technique was used to estimate relative concentrations of metabolites of xenobiotics in bile of 103 English sole (Parophrys vetulus) from both polluted and minimally polluted (reference) sites in Puget Sound, WA. Fish from polluted sites had concentrations of xenobiotics in bile with naphthalene-, phenanthrene- and benzo[a]pyrene-like fluorescence that averaged 9, 14 and 19 times, respectively, those of fish from reference sites. Within a polluted site, fish with liver lesions had significantly higher bile concentrations of xenobiotics with benzo[a]pyrene-like fluorescence than did fish without liver lesions. Individual metabolites of fluorene, phenanthrene, anthracene, biphenyl and dimethylnaphthalene were determined by g.l.c.-mass spectrometry in extracts of hydrolysed bile of three English sole from polluted waterways; concentrations ranged from 90 to 19000 ng/g, wet wt. Other xenobiotics were tentatively identified, but not quantified.

Topics: Animals; Benzo(a)pyrene; Bile; Chemical and Drug Induced Liver Injury; Chromatography, High Pressure Liquid; Fish Diseases; Fishes; Hydrocarbons; Liver Diseases; Naphthalenes; Phenanthrenes; Spectrometry, Fluorescence; Water Pollutants; Water Pollutants, Chemical