Triptolide Induces hepatotoxicity via inhibition of CYP450s in Rat liver microsomes
Autor: | Fuqiong Zhou, Xueping Zhou, Yan Lu, Huaxu Zhu, Zhe Feng, Weina Zhu, Tong Xie, Jie Ruan, Yajie Zhang |
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Jazyk: | angličtina |
Předmět: |
0301 basic medicine
Male CYP3A Metabolite CYP450s Pharmacology Isozyme Rats Sprague-Dawley 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Cytochrome P-450 Enzyme System In vivo Microsomes Animals Cytochrome P-450 Enzyme Inhibitors biology Triptolide Hepatotoxicity Cytochrome P450 General Medicine CYP2E1 Phenanthrenes Rats Isoenzymes Kinetics 030104 developmental biology chemistry Liver Complementary and alternative medicine 030220 oncology & carcinogenesis Microsome biology.protein Microsomes Liver Epoxy Compounds Diterpenes Research Article |
Zdroj: | BMC Complementary and Alternative Medicine |
ISSN: | 1472-6882 |
DOI: | 10.1186/s12906-016-1504-3 |
Popis: | Background Triptolide (TP), an active constituent of Tripterygium wilfordii, possesses numerous pharmacological activities. However, its effects on cytochrome P450 enzymes (CYP450s) in rats remain unexplored. Methods In this study, the effects of triptolide on the six main CYP450 isoforms (1A2, 2C9, 2C19, 2D6, 2E1, and 3A) were investigated both in vivo and in vitro. We monitored the body weight, survival proportions, liver index, changes in pathology, and biochemical index upon TP administration, in vivo. Using a cocktail probe of CYP450 isoform-specific substrates and their metabolites, we then carried out in vitro enzymatic studies in liver microsomal incubation systems via ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Finally, we verified our results at the messenger ribonucleic acid (mRNA) and protein level through quantitative real-time polymerase chain reaction (RT-qPCR), western blotting, and immunohistochemical detection. Results The in vivo toxicity study confirmed that Sprague-Dawley (SD) rats exhibited dose-dependent hepatotoxicity after intragastric administration of TP [200, 400, and 600 μg/(kg.day)] for 28 days. In case of the CYP450 isoforms 3A, 2C9, 2C19, and 2E1, the in vitro metabolic study demonstrated a decrease in the substrate metabolic rate, metabolite production rate, and Vmax, with an increase in the Km value, compared with that observed in the control group. Additionally, a TP dose-dependent decrease in the mRNA levels was observed in the four major isoforms of CYP3A subfamily (3A1/3A23, 3A2, 3A9, and 3A62) and CYP2C9. A similar effect was also observed with respect to the protein levels of CYP2C19 and CYP2E1. Conclusions This study suggests that TP can cause hepatotoxicity by reducing the substrate affinity, activity, and expression at the transcriptional and protein levels of the CYP450 isoforms 3A, 2C9, 2C19, and 2E1. TP also has the potential to cause pharmacokinetic drug interactions when co-administered with drugs metabolized by these four isoforms. However, further clinical studies are needed to evaluate the significance of this interaction. Electronic supplementary material The online version of this article (doi:10.1186/s12906-016-1504-3) contains supplementary material, which is available to authorized users. |
Databáze: | OpenAIRE |
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