附子毒性生物碱的肝脏摄取机制  

Uptake Mechanism of Toxic Alkaloids of Fuzi in Liver

作　　者：从娇娇 张雷 覃晓慧 张峻颖[4] 吴春勇[1,2] CONG Jiaojiao;ZHANG Lei;QIN Xiaohui;ZHANG Junying;WU Chunyong(Key Laboratory of Drug Quality Control and Pharmacovigilance, Ministry of Education, Nanjing 210009,China;Department of Pharmaceutical Analysis, China Pharmaceutical University, Nanjing 210009, China;Shandong Institute for Food andDrug Control, Shandong 250101, China;Department of Pharmaceutics of Traditional Chinese Medicine, China Pharmaceutical University,Nanjing 211198, China)

机构地区：[1]中国药科大学药物质量与安全预警教育部重点实验室,南京210009 [2]中国药科大学药物分析系,南京210009 [3]山东省食品药品检验研究院,济南250101 [4]中国药科大学中药制剂教研室,南京211198

出　　处：《医药导报》2019年第6期711-717,共7页Herald of Medicine

基　　金：国家自然科学基金资助项目(81473357;81673681);中央高校基本科研业务费项目(2015PT062);江苏高校优势学科建设工程资助项目;双一流建设项目(CPU2018GY34)

摘　　要：目的探讨附子主要毒性生物碱成分乌头碱、新乌头碱和次乌头碱的肝脏摄取转运机制。方法以HepG_2细胞为体外模型,进行细胞摄取研究,并用液相色谱-质谱串联(LC-MS/MS)法测定细胞内的药物浓度。结果 HepG_2细胞对乌头碱、新乌头碱和次乌头碱的摄取具有温度、浓度和pH依赖性,化学解耦联剂FCCP和能量抑制剂叠氮化钠可显著抑制乌头碱、新乌头碱和次乌头碱的细胞摄取。有机阳离子药物如奎尼丁、维拉帕米、吡拉明、苯海拉明及金刚烷胺可显著抑制3种生物碱的摄取,但有机阳离子转运蛋白(OCTs)、有机阳离子/肉毒碱转运蛋白(OCTNs)、多药及毒素外排转运蛋白(MATEs)和质膜单胺转运蛋白(PMAT)的典型底物或抑制剂对附子生物碱的细胞摄取没有抑制作用。结论附子毒性生物碱进入肝细胞以主动转运为主,新型有机阳离子/质子逆向转运蛋白可能是其进入肝脏的分子基础。Objective To characterize the liver uptake of the main toxic alkaloids of Fuzi including aconitine, mesaconitine and hypaconitine. Methods Celler uptake was studied by using HepG2 cells as an in vitro model, and the drug concentration was determined by liquid chromatography-mass spectrometry (LC-MS/MS). Results Aconitine, mesaconitine and hypaconitine were transported into HepG2 cells in a temperature-, concentration- and pH-dependent manner, which were significantly reduced by FCCP (a protonophore) and sodium azide (a metabolic inhibitor).The uptakes of Fuzi alkaloids were greatly inhibited by various cationic drugs such as quinidine, verapamil, pyrilamine, diphenhydramine and amantadine.However, the prototypical substrates or inhibitors of well-characterized organic cation transporter (OCTs), organic cation/carnitine transporters (OCTNs), multidrug and toxin extrusion transporters (MATEs) and plasma membrane monoamine transporter (PMAT) didn’t inhibit the cellular uptake of each Fuzi alkaloid. Conclusion Toxic alkaloids of Fuzi are mainly actively transported into liver cells, and the novel organic cation/proton antiporters may be the molecular basis for their entry into liver.

关 键 词：附子 生物碱 毒性 细胞摄取 有机阳离子/质子逆向转运蛋白 HEPG2细胞 

分 类 号：R285[医药卫生—中药学]