The contribution of human OCT1, OCT3, and CYP3A4 to nitidine chloride-induced hepatocellular toxicity.
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 NC-induced 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 NC-induced toxicity. In addition, P450 enzymes, such as CYP3A4, mediated the metabolism of NC, and NC-induced toxicity in MDCK-hOCT1/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.
Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences (L.L., M.T., X.Y., S.S., H.Z., S.Z., H.J.) and Center of Analysis and Measurement (X.W.), Zhejiang University, Hangzhou, Zhejiang, China.
Liping
L
Li
Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences (L.L., M.T., X.Y., S.S., H.Z., S.Z., H.J.) and Center of Analysis and Measurement (X.W.), Zhejiang University, Hangzhou, Zhejiang, China.
Meijuan
M
Tu
Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences (L.L., M.T., X.Y., S.S., H.Z., S.Z., H.J.) and Center of Analysis and Measurement (X.W.), Zhejiang University, Hangzhou, Zhejiang, China.
Xi
X
Yang
Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences (L.L., M.T., X.Y., S.S., H.Z., S.Z., H.J.) and Center of Analysis and Measurement (X.W.), Zhejiang University, Hangzhou, Zhejiang, China.
Siyuan
S
Sun
Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences (L.L., M.T., X.Y., S.S., H.Z., S.Z., H.J.) and Center of Analysis and Measurement (X.W.), Zhejiang University, Hangzhou, Zhejiang, China.
Xiaodan
X
Wu
Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences (L.L., M.T., X.Y., S.S., H.Z., S.Z., H.J.) and Center of Analysis and Measurement (X.W.), Zhejiang University, Hangzhou, Zhejiang, China.
Hui
H
Zhou
Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences (L.L., M.T., X.Y., S.S., H.Z., S.Z., H.J.) and Center of Analysis and Measurement (X.W.), Zhejiang University, Hangzhou, Zhejiang, China.
Su
S
Zeng
Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences (L.L., M.T., X.Y., S.S., H.Z., S.Z., H.J.) and Center of Analysis and Measurement (X.W.), Zhejiang University, Hangzhou, Zhejiang, China hdjiang@zju.edu.cn.
Huidi
H
Jiang
D000818:Animals; D053119:Benzophenanthridines / Q000633:toxicity*; D002460:Cell Line; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism*; D004285:Dogs; D006801:Humans; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000378:metabolism*; D027702:Organic Cation Transporter 1 / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
D053119:Benzophenanthridines; D050814:Octamer Transcription Factor-3; D027702:Organic Cation Transporter 1; C494173:POU5F1 protein, human; C013615:nitidine; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human