Organic cation transporter 1 mediates the uptake of monocrotaline and plays an important role in its hepatotoxicity

Tu, Meijuan; Sun, Siyuan; Wang, Kai; Peng, Xueying; Wang, Ruihan; Li, Liping; Zeng, Su; Zhou, Hui; Jiang, Huidi

doi:10.1016/j.tox.2013.06.009

Cited by 54 publications

(59 citation statements)

References 37 publications

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“…As shown in Fig. 2A, NC inhibited the uptake of MPP + in a concentration-dependent manner, with IC 50 values of 1.09 6 0.077 mM and 0.638 6 0.12 mM, respectively, while the IC 50 values of two known inhibitors of OCT1/OCT3, quinidine and (+)-THP (Tu et al, 2013), were 2.47 6 0.74 mM and 12.4 6 1.7 mM for hOCT1 and 1.52 6 0.28 mM and 9.42 6 0.37 mM for hOCT3, respectively (Fig. 2, B and C).…”

Section: Resultsmentioning

confidence: 81%

“…The MDCK cell line was obtained from Peking Union Medical College (Beijing, China). MDCK cells stably transfected with plasmid pcDNA3.1(+) vector containing human OCT1 cDNA (MDCK-hOCT1), human OCT3 cDNA (MDCK-hOCT3), human MATE1 cDNA (MDCK-hMATE1), and empty vector (mock cells) were constructed in our laboratory (Tu et al, 2013;Sun et al, 2014). The activity of hOCT1 and hOCT3 in the stably transfected cells was validated by model substrates, such as ASP + and MPP + , with or without quinidine, an OCT1 and OCT3 inhibitor (Hasannejad et al, 2004;Ahlin et al, 2008).…”

Section: Methodsmentioning

confidence: 99%

“…MDCK-hOCT1, MDCK-hOCT3, and mock cells were seeded in 24-well plates at a density of 2 Â 10 5 /well. Cells were cultured for 3 days and washed twice with Hanks' balanced salt solution (HBSS); then the uptake studies were performed as in our previous method (Tu et al, 2013). Briefly, the cells were preincubated with HBSS (37°C, 10 minutes), and then 200 ml of HBSS containing MPP + or NC in the absence or presence of different inhibitors was added to initiate the uptake.…”

Section: Methodsmentioning

confidence: 99%

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The Contribution of Human OCT1, OCT3, and CYP3A4 to Nitidine Chloride–Induced Hepatocellular Toxicity

Yang

et al. 2014

Drug Metab Dispos

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Nitidine chloride (NC), a quaternary ammonium alkaloid, has numerous pharmacological effects, such as anticancer activity. However, it was found that NC also has hepatocellular toxicity. Because organic cation transporters 1 and 3 (OCT1 and OCT3) might mediate the influx of NC into hepatocytes, multidrug and toxin extrusion 1 (MATE1) probably mediates the efflux of NC from hepatocytes, while cytochrome P450 (P450) enzymes might contribute to NC metabolism, the present study was to evaluate the contribution of OCT1, OCT3, MATE1, and P450 enzymes to NCinduced hepatocellular toxicity. Our results showed that the uptake of NC in Madin-Darby canine kidney (MDCK) cells expressing human (h) OCT1 and OCT3 (MDCK-hOCT1 and MDCK-hOCT3) was significantly higher than that in mock cells; the hOCT1-and hOCT3-mediated uptake followed typical Michaelis-Menten kinetics.Meanwhile, NC was also a substrate of hMATE1, although its transport capacity was much lower than that of OCT1 NC-induced cytotoxicity in MDCK-hOCT1 or MDCK-hOCT3 cells was obviously higher than that in mock cells. Quinidine and (+)-tetrahydropalmatine [(+)-THP], OCT1 and OCT3 inhibitors, significantly reduced the uptake of NC in MDCK-hOCT1 cells, MDCK-hOCT3 cells, and rat primary hepatocytes, but only (+)-THP markedly attenuated the NCinduced toxicity. In addition, P450 enzymes, such as CYP3A4, mediated the metabolism of NC, and NC-induced toxicity in MDCKhOCT1/hCYP3A4 cells was lower than that in MDCK-hOCT1 cells. Our results indicated that NC is a substrate of hOCT1, hOCT3, and CYP3A4; that OCT1 and OCT3 mediate the uptake of NC in hepatocytes and subsequently cause hepatotoxicity; and that NC-induced toxicity could be attenuated by CYP3A4-mediated metabolism.

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Section: Resultsmentioning

confidence: 81%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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The Contribution of Human OCT1, OCT3, and CYP3A4 to Nitidine Chloride–Induced Hepatocellular Toxicity

Yang

et al. 2014

Drug Metab Dispos

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“…MDCK cells stably expressing hOCT2, hOCTN1, hOCTN2, hMATE1, hMATE2-K, hMDR1, hMRP2, or mock-transfected cells were seeded in 24-well plates (Costar Corning Inc., Corning, NY, USA) at a density of 2 ϫ 10 5 /well. On day 3 after seeding, the accumulation studies were performed according to methods developed in our laboratory (14)(15)(16)21). The effectiveness of the transgenic cell models was verified using probe substrates including MPP ϩ (1 M) for hOCT2 and hMATE1/2-K, L-ergothioneine (3 M) for hOCTN1, mildronate (2 M) for hOCTN2, and Rho123 (4 M) for hMDR1 and calcein AM (1 M) for hMRP2.…”

mentioning

confidence: 99%

“…The concentrations of MPP ϩ , L-ergothioneine, and mildronate in the samples were determined by an Agilent 1290/6460 LC-MS with a triple-quadrupole mass spectrometer (Agilent Technologies, Santa Clara, CA) according to the method described previously (21,23). For ETV determination, 80 l of the cell lysate was mixed with 240 l acetonitrile containing the internal standard (5 ng/ml MPP ϩ ) for 3 min, and then the mixture was centrifuged for 15 min at 16,000 ϫ g and the supernatant was analyzed by LC-MS/MS.…”

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confidence: 99%

Multiple Drug Transporters Are Involved in Renal Secretion of Entecavir

Yang

Zhou

et al. 2016

Antimicrob Agents Chemother

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Entecavir (ETV) is a first-line antiviral agent for the treatment of chronic hepatitis B virus infection. Renal excretion is the major elimination path of ETV, in which tubular secretion plays the key role. However, the secretion mechanism has not been clarified. We speculated that renal transporters mediated the secretion of ETV. Therefore, the aim of our study was to elucidate which transporters contribute to the renal disposition of ETV. Our results revealed that ETV (50 M) remarkably reduced the accumulation of probe substrates in MDCK cells stably expressing human multidrug and toxin efflux extrusion proteins (hMATE1/2-K), organic cation transporter 2 (hOCT2), and carnitine/organic cation transporters (hOCTNs) and increased the substrate accumulation in cells transfected with multidrug resistance-associated protein 2 (hMRP2) or multidrug resistance protein 1 (hMDR1). Moreover, ETV was proved to be a substrate of the above-described transporters. In transwell studies, the transport of ETV in MDCK-hOCT2-hMATE1 showed a distinct directionality from BL (hOCT2) to AP (hMATE1), and the cellular accumulation of ETV in cells expressing hMATE1 was dramatically lower than that of the mock-treated cells. The accumulation of ETV in mouse primary renal tubular cells was obviously affected by inhibitors of organic anion transporter 1/3 (Oat1/3), Oct2, Octn1/2, and Mrp2. Therefore, the renal uptake of ETV is likely mediated by OAT1/3 and OCT2 while the efflux is mediated by MATEs, MDR1, and MRP2, and OCTN1/2 may participate in both renal secretion and reabsorption. C hronic hepatitis B virus (HBV) infection, with a high rate of morbidity, is one of the most important health problems worldwide. It is a major risk factor for cirrhosis and liver cancer (1). Entecavir (ETV) is a novel and highly selective deoxyguanosine analog with a high antiviral efficacy and a high genetic barrier to viral resistance (2). Since ETV was approved in 2005 by the U.S. FDA, it has been unanimously recommended as the fist-line antiviral agent for treatment of chronic HBV infection by global hepatology scientific societies, such as the American Association for the Study of Liver Diseases and the European Association for the Study of the Liver (3, 4).It was reported that about 73% of orally dosed ETV was eliminated in urine in an unchanged form. The renal clearance of ETV is independent of the dose and ranged from 360 to 471 ml/min in healthy volunteers, which is much greater than the glomerular filtration rate (GFR; 120 to 130 ml/min for normal humans) (5, 6), suggesting that tubular secretion accounts for the major part of the urinary excretion of ETV. Therefore, it is speculated that transporters expressed on the proximal tubular cells are involved in the process of tubular secretion of ETV. Furthermore, ETV is a hydrophilic weak base with a pK a value of 10.5 (Ͼ99% of ETV is positively charged at pH 7.4) and a logD value of Ϫ1.1 (pH 4 to 10), which implies that ETV is unlikely to penetrate the cell membrane by passive diffusion. Thus, transpor...

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Drug Routes of Excretion

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Organic cation transporter 1 mediates the uptake of monocrotaline and plays an important role in its hepatotoxicity

Cited by 54 publications

References 37 publications

The Contribution of Human OCT1, OCT3, and CYP3A4 to Nitidine Chloride–Induced Hepatocellular Toxicity

The Contribution of Human OCT1, OCT3, and CYP3A4 to Nitidine Chloride–Induced Hepatocellular Toxicity

Multiple Drug Transporters Are Involved in Renal Secretion of Entecavir

Drug Routes of Excretion

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