Utilizing single- and double-transfected cell models expressing human organic anion transporter 1 and human cytochrome P450 1A2 to investigate the interactions with ingredients of herbal medicines

Abstract

1. Cell models expressing human drug transporters and enzymes are useful tools to understand the process of drug disposition in vitro. However, no study on transfected cells stably co-expressing human organic anion transporter 1 (hOAT1) and/or human cytochrome P450 1A2 (hCYP1A2) is available. In this study, cell models stably expressing hOAT1 and/or hCYP1A2 were established, and were used to investigate the interactions of ingredients of herbal medicines (IHMs) with hOAT1 and/or hCYP1A2.

2. The MDCK cells were stable transfected with recombinant plasmids expressing hOAT1 and/or hCYP1A2. Cellular uptake assay and CYP450 activity assay showed that the transfected cells were available. A marked high expression of hOAT1 and hCYP1A2 mRNA was also validated by quantitative RT-PCR. Totally 6 IHMs which significantly inhibited the activity of hOAT1 were screened out by employing hOAT1 expressing cells. The contribution of hOAT1 and hCYP1A2 to the toxicity of aristolochic acid I (AAI) was further determined. Compared to mock cells, all transfected cells showed a decrease in viability after being treated with AAI.

3. A method to establish transfected cell expressing drug metabolism enzymes and/or transporters was provided in our study. Three IHMs (dihydrotanshinone I, cryptotanshinone, and tanshinone I) were confirmed as novel inhibitors of hOAT1. Furthermore, a synergistic effect of hOAT1 and hCYP1A2 on AAI-induced toxicity was also observed in this investigation.

Keywords:

• Aristolochic acid I
• cytotoxicity
• hCYP1A2
• hOAT1
• ingredients of herbal medicines
• stable-transfected cell models

Declaration of interest

This project was supported by International Science & Technology Cooperation Program of China (2014DFE30050), Ministry of Science and Technology of China (2012ZX09506001-004), the Program for Zhejiang Leading Team of S&T Innovation Team (2011R50014), and the Fundamental Research Funds for the Central Universities of China Ministry of Education (2016XZZX001-08). The authors report no declarations of interest.