Time-Resolved Effect of Interferon-Alpha 2a on Activities of Nuclear Factor Kappa B, Pregnane X Receptor and on Drug Disposition Genes

Theile, Dirk; Wagner, Lelia; Bay, Cindy; Haefeli, Walter E.; Weiß, Johanna

doi:10.3390/pharmaceutics13060808

Cited by 3 publications

(2 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cells were subsequently incubated for 20 min at room temperature. Luminescence was recorded immediately and normalized to cell number using crystal violet staining (Theile et al 2021 ). CYP3A4 activity was normalized to untreated controls.…”

Section: Methodsmentioning

confidence: 99%

Comprehensive in vitro analysis evaluating the variable drug–drug interaction risk of rifampicin compared to rifabutin

et al. 2023

Self Cite

View full text Add to dashboard Cite

Compared to rifampicin (600 mg/day), standard doses of rifabutin (300 mg/day) have a lower risk of drug–drug interactions due to induction of cytochrome P450 3A4 (CYP3A4) or P-glycoprotein (Pgp/ABCB1) mediated by the pregnane X receptor (PXR). However, clinical comparisons with equal rifamycin doses or in vitro experiments respecting actual intracellular concentrations are lacking. Thus, the genuine pharmacological differences and the potential molecular mechanisms of the discordant perpetrator effects are unknown. Consequently, the cellular uptake kinetics (mass spectrometry), PXR activation (luciferase reporter gene assays), and impact on CYP3A4 and Pgp/ABCB1 expression and activity (polymerase chain reaction, enzymatic assays, flow cytometry) were evaluated in LS180 cells after treatment with different rifampicin or rifabutin concentrations for variable exposure times and eventually normalized to actual intracellular concentrations. In addition, inhibitory effects on CYP3A4 and Pgp activities were investigated. While rifampicin is poorly taken up by LS180 cells, it strongly activates PXR and leads to enhanced expression and activity of CYP3A4 and Pgp. In contrast, rifabutin is a significantly less potent and less efficient PXR activator and gene inducer, despite sixfold to eightfold higher intracellular accumulation. Finally, rifabutin is a potent inhibitor of Pgp (IC50 = 0.3 µM) compared to rifampicin (IC50 = 12.9 µM). Together, rifampicin and rifabutin significantly differ by their effects on the regulation and function of CYP3A4 and Pgp, even when controlled for intracellular concentrations. Rifabutin’s concurrent Pgp inhibitory action might partly compensate the inducing effects, explaining its weaker clinical perpetrator characteristics.

show abstract

Section: Methodsmentioning

confidence: 99%

Comprehensive in vitro analysis evaluating the variable drug–drug interaction risk of rifampicin compared to rifabutin

et al. 2023

Self Cite

View full text Add to dashboard Cite

show abstract

“…Other factors also affect drug pharmacokinetics through PXR. Interferon-alpha has been suggested to cause pharmacokinetic drug interactions by decreasing the expression of drug disposition genes by influencing NF-κB and PXR activities (Theile et al, 2021). Besides PXR, CAR can influence drug pharmacokinetics through the regulation of CYP1A, UGTs, and SULTs (Fu et al, 2020).…”

mentioning

confidence: 99%

The Function of Xenobiotic Receptors in Metabolic Diseases

Zhang

Jia

et al. 2022

Drug Metab Dispos

View full text Add to dashboard Cite

Metabolic diseases are a series of metabolic disorders that include obesity, diabetes, insulin resistance, hypertension, and hyperlipidemia. The increased prevalence of metabolic diseases has resulted in higher mortality and mobility rates over the past decades, and this has led to extensive research focusing on the underlying mechanisms. Xenobiotic receptors (XRs) are a series of xenobioticsensing nuclear receptors that regulate their downstream target genes expression, thus defending the body from xenobiotic and endotoxin attacks. XR activation is associated with the development of a number of metabolic diseases such as obesity, nonalcoholic fatty liver disease (NAFLD), type 2 diabetes (T2D), and cardiovascular diseases, thus suggesting an important role for XRs in modulating metabolic diseases.However, the regulatory mechanism of XRs in the context of metabolic disorders under different nutrient conditions is complex and remains controversial. This review summarizes the effects of XRs on different metabolic components (cholesterol, lipids, glucose, and bile acids) in different tissues during metabolic diseases. As chronic inflammation plays a critical role in the initiation and progression of metabolic diseases, we also discuss the impact of XRs on inflammation to comprehensively recognize the role of XRs in metabolic diseases. This will provide new ideas for treating metabolic diseases by targeting XRs.

show abstract

Crystal Violet Staining is a Reliable Alternative to Bicinchoninic Acid Assay-Based Normalization

2022

View full text Add to dashboard Cite

Experimental data with cells often require normalization. The frequently used bicinchoninic acid (BCA) assay, in fact, indicates protein content but is influenced by incubation time, pH etc. A simple, rapid and reliable alternative is desirable. Crystal violet stains nucleic acids and proteins and was used to reflect the cell number in 96-well plates. Calibration curves and comparison with BCA confirmed excellent goodness of fit (R2: 0.98), conformity (nonsignificant difference of BCA to crystal violet) and reliability of this staining methodology. Crystal violet staining can be used to normalize experimental data to the number of adherent cells present in cell culture plates.

show abstract

Time-Resolved Effect of Interferon-Alpha 2a on Activities of Nuclear Factor Kappa B, Pregnane X Receptor and on Drug Disposition Genes

Cited by 3 publications

References 43 publications

Comprehensive in vitro analysis evaluating the variable drug–drug interaction risk of rifampicin compared to rifabutin

Comprehensive in vitro analysis evaluating the variable drug–drug interaction risk of rifampicin compared to rifabutin

The Function of Xenobiotic Receptors in Metabolic Diseases

Crystal Violet Staining is a Reliable Alternative to Bicinchoninic Acid Assay-Based Normalization

Contact Info

Product

Resources

About