Min Hongping, Niu Miaomiao, Zhang Weilin, Yan Jia, Li Jiachang, Tan Xiying, Li Bo, Su Mengxiang, Di Bin, Yan Fang
Key Laboratory on Protein Chemistry and Structural Biology, China Pharmaceutical University, Nanjing, China.
Key Laboratory of Drug Quality Control and Pharmacovigilance (China Pharmaceutical University), Ministry of Education, Nanjing, China.
PLoS One. 2017 Nov 9;12(11):e0187971. doi: 10.1371/journal.pone.0187971. eCollection 2017.
Development of multidrug resistance (MDR) is a continuous clinical challenge partially due to the overexpression of P-glycoprotein (P-gp) for chronic myelogenous leukemia (CML) patients. Herein, we evaluated the inhibitory potency of emodin, a natural anthraquinone derivative isolated from Rheum palmatum L, on P-gp in P-gp positive K562/ADM cells. Competition experiments combined with molecular docking analysis were utilized to investigate the binding modes between emodin and binding sites of P-gp. Emodin reversed adriamycin resistance in K562/ADM cells accompanied with the decrease of P-gp protein expression, further increasing the uptake of rhodamine123 in both K562/ADM and Caco-2 cells, indicating the inhibition of P-gp efflux function. Moreover, when incubated with emodin under different conditions where P-gp was inhibited, K562/ADM cells displayed increasing intracellular uptake of emodin, suggesting that emodin may be the potential substrate of P-gp. Importantly, rhodamine 123 could increase the Kintrinsic (Ki) value of emodin linearly, whereas, verapamil could not, implying that emodin competitively bound to the R site of P-gp and noncompetition existed between emodin and verapamil at the M site, in a good accordance with the results of molecular docking that emodin bound to the R site of P-gp with higher affinity. Based on our results, we suggest that emodin might be used to modulate P-gp function and expression.
多药耐药性(MDR)的产生是一个持续的临床挑战,部分原因是慢性粒细胞白血病(CML)患者中P-糖蛋白(P-gp)的过度表达。在此,我们评估了从掌叶大黄中分离出的天然蒽醌衍生物大黄素对P-gp阳性K562/ADM细胞中P-gp的抑制效力。利用竞争实验结合分子对接分析来研究大黄素与P-gp结合位点之间的结合模式。大黄素逆转了K562/ADM细胞中的阿霉素耐药性,同时P-gp蛋白表达降低,进一步增加了罗丹明123在K562/ADM和Caco-2细胞中的摄取,表明对P-gp外排功能的抑制。此外,在不同条件下用大黄素孵育抑制P-gp时,K562/ADM细胞对大黄素的细胞内摄取增加,表明大黄素可能是P-gp的潜在底物。重要的是,罗丹明123可使大黄素的内在解离常数(Ki)值呈线性增加,而维拉帕米则不能,这意味着大黄素竞争性地结合到P-gp的R位点,且大黄素与维拉帕米在M位点不存在竞争,这与分子对接结果很好地相符,即大黄素以更高的亲和力结合到P-gp的R位点。基于我们的结果,我们认为大黄素可能用于调节P-gp的功能和表达。