RESUMO
Cytochrome P450, which is expressed in humans and other animals, is a superfamily of drug-metabolizing enzymes that play important roles in the metabolism of endogenous and xenobiotic substrates via oxidation, peroxidation and reduction. Of endogenous substrates, interleukin (IL)-6 is a crucial cytokine involved in inflammation in the liver. The present study aims to elucidate the mechanisms through which IL-6 modulates cytochrome P450 expression. CYP2C33 expression was found to be increased in HepLi cells and primary porcine hepatocytes treated with IL-6 in a concentration-dependent manner. IL-6 treatment also increased the expression of the transcriptional regulators, constitutive androstane receptor (CAR) and pregnane X receptor. Overexpression of CAR promoted CYP2C33 expression at the mRNA and protein levels, whereas knockdown of CAR by small interfering RNA reduced CYP2C33 expression. Luciferase assays showed that IL-6 treatment of HepLi cells and primary porcine hepatocytes increased CYP2C33 promoter activity. Co-immunoprecipitation and western blotting demonstrated that CAR and RXR could form heterodimers. IL-6 affects CYP2C33 expression through CAR/retinoid X receptor (RXR) heterodimers.
Assuntos
Sistema Enzimático do Citocromo P-450/metabolismo , Interleucina-6/farmacologia , Receptor de Pregnano X/fisiologia , Receptores Citoplasmáticos e Nucleares/fisiologia , Animais , Linhagem Celular , Receptor Constitutivo de Androstano , Sistema Enzimático do Citocromo P-450/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Hepatócitos/metabolismo , Inativação Metabólica , Interleucina-6/metabolismo , Interleucina-6/fisiologia , Receptor de Pregnano X/genética , Receptor de Pregnano X/metabolismo , RNA Mensageiro/metabolismo , Receptores Citoplasmáticos e Nucleares/genética , Receptores Citoplasmáticos e Nucleares/metabolismo , Suínos , Xenobióticos/metabolismoRESUMO
The objective of this study was to provide evidence of the validity of utilizing pigs as a model to study the regulation of human CYP3A4, with special emphasis on drug-drug interactions. We determined the mRNA expression and distribution of CYP3A and metabolic nuclear receptors in different tissues isolated from landrace pigs. Our results showed that CYP3A and metabolic nuclear receptor mRNAs were most highly expressed in liver tissues. The expression of the metabolic nuclear receptor pregnane X receptor (PXR) had a significant correlation with expression of CYP3A29, an analog of human CYP3A4. The correlation between their transcriptional levels was further demonstrated using LPS and TNF-α. The mRNA and protein expression of CYP3A29 and PXR in HepLi cells was significantly reduced by LPS and TNF-α treatment. CYP3A29 promoter activity was dramatically elevated by PXR over expression, whereas LPS and TNF-α treatment inhibited the enhanced CYP3A29 promoter activity that was induced by PXR; presumably through inhibition of PXR promoter activity. Furthermore, the inhibition of CYP3A29 promoter activity by LPS and TNF-α treatment was blocked by knockdown of PXR or retinoid X receptor (RXR). These data suggest high similarity in the regulation mechanism of pig CYP3A29 and human CYP3A4. Our research provided a significant evaluation to determine whether pigs are suitable as an experimental animal model.