Multidrug resistance-associated protein 2 is involved in the efflux of Aconitum alkaloids determined by MRP2-MDCKII cells.
OBJECTIVE
Aconitum alkaloids mainly contain highly toxic aconitine (AC), mesaconitine (MA), and hypaconitine (HA) and less toxic benzoylaconine (BAC), benzoylmesaconine (BMA), benzoylhypaconine (BHA), aconine, mesaconine, and hypaconine. The efflux transporters including P-glycoprotein (P-gp), breast cancer resistance protein (BCRP) and multidrug resistance-associated protein 2 (MRP2) can efflux toxicants to prevent poisoning. Our previous publication has proved that P-gp and BCRP contributed to the efflux of AC, MA and HA, which is demonstrated in the human colonic adenocarcinoma cell lines (Caco-2 cells), Mardin-Darby canine kidney cell lines transfected with MDR1 or BCRP (MDR1-MDCKII and BCRP-MDCKII cells). However, the role of MRP2 remains uncertain.


METHODS
The MRP2-MDCKII cells were used to determine the efflux ratios (Er) and intracellular amounts of Aconitum alkaloids. In addition, the importance of MRP2 was further investigated with or without the MRP2 inhibitor, LTC4.


RESULTS
The Er values of AC, MA, HA, BAC, BMA and BHA in MRP2-MDCKII cells (6.4 ± 0.3, 5.9 ± 0.5, 2.2 ± 0.2, 1.6 ± 0.3, 1.7 ± 0.2 and 1.9 ± 0.2 respectively) were significantly higher than those in MDCKII cells, which were close to 1. In the presence of LTC4, the Er values of AC, MA, HA, BAC, BMA and BHA were reduced to approximately 1 and their intracellular amounts were also significantly increased in MRP2-MDCKII cells.


CONCLUSIONS
MRP2 was involved in the efflux of AC, MA, HA, BAC, BMA and BHA, which would be useful for the safe application of these components or their herbs.
International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China; Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong 510515, China.
Peimin
P
Dai
International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China.
Lijun
L
Zhu
International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China.
Xiaoshan
X
Yang
International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China.
Min
M
Zhao
International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China; Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong 510515, China.
Jian
J
Shi
International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China; Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong 510515, China.
Ying
Y
Wang
International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China. Electronic address: lllu@gzucm.edu.cn.
Linlin
L
Lu
International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China; Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong 510515, China. Electronic address: liuzq@gzucm.edu.cn.
Zhongqiu
Z
Liu
D018435:ATP Binding Cassette Transporter, Subfamily B / Q000378:metabolism; D031841:Aconitum / Q000737:chemistry*; D000470:Alkaloids / Q000378:metabolism*; D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D018938:Caco-2 Cells; D004285:Dogs; D006801:Humans; D017997:Leukotriene C4 / Q000494:pharmacology; D061985:Madin Darby Canine Kidney Cells; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014162:Transfection
D018435:ATP Binding Cassette Transporter, Subfamily B; D000470:Alkaloids; D017997:Leukotriene C4; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; C513055:ABCB1 protein, human; C000715087:ABCC2 protein, human
Aconitine (PubChem CID: 245005); Aconitum alkaloids; Efflux transporters; MRP2-MDCKII cell; Multidrug resistance-associated protein 2; aconine (PubChem CID: 11969787); benzoylaconine (PubChem CID: 121312); benzoylhypaconine (PubChem CID: 24832662); benzoylmesaconine (PubChem CID: 24832659); hypaconine (PubChem CID: 78358526); hypaconitine (PubChem CID: 4526188); mesaconine (PubChem CID: 24832657); mesaconitine (PubChem CID: 416228)