title: 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.
authors.affiliation: a Institute of Drug Metabolism and Pharmaceutical Analysis, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , China.
authors.forename: Zhiyuan
authors.initials: Z
authors.lastname: Qin
authors.affiliation: a Institute of Drug Metabolism and Pharmaceutical Analysis, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhao
authors.affiliation: a Institute of Drug Metabolism and Pharmaceutical Analysis, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , China.
authors.forename: Haihong
authors.initials: H
authors.lastname: Hu
authors.affiliation: a Institute of Drug Metabolism and Pharmaceutical Analysis, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , China.
authors.forename: Huidi
authors.initials: H
authors.lastname: Jiang
authors.affiliation: a Institute of Drug Metabolism and Pharmaceutical Analysis, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , China.
authors.forename: Lushan
authors.initials: L
authors.lastname: Yu
authors.affiliation: a Institute of Drug Metabolism and Pharmaceutical Analysis, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University , Hangzhou , China.
authors.forename: Su
authors.initials: S
authors.lastname: Zeng
mesh_terms: D000818:Animals; D001692:Biological Transport; D019388:Cytochrome P-450 CYP1A2 / Q000378:metabolism*; D004285:Dogs; D041743:Herb-Drug Interactions; D029001:Herbal Medicine; D006801:Humans; D061985:Madin Darby Canine Kidney Cells; D008954:Models, Biological; D027362:Organic Anion Transport Protein 1 / Q000378:metabolism*; D014162:Transfection
chemical_list: D027362:Organic Anion Transport Protein 1; D019388:Cytochrome P-450 CYP1A2
keywords: Aristolochic acid I; cytotoxicity; hCYP1A2; hOAT1; ingredients of herbal medicines; stable-transfected cell models