丹参酮成分对胆汁酸体外羟化代谢的抑制研究  

Inhibitory effects of tanshinones on hydroxylation metabolism of bile acids in vitro

作　　者：李玥 王倩 苏慧宗 谭波 金静怡 宋国超 严东明 裘福荣 LI Yue;WANG Qian;SU Huizong;TAN Bo;JIN Jingyi;SONG Guochao;YAN Dongming;QIU Furong(Clinical Laboratory of Pharmacokinetics,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院临床药代动力学实验室,上海201203

出　　处：《上海中医药杂志》2020年第S01期177-182,共6页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目81874346);上海市中医临床重点实验室项目(14DZ2273200);上海中医药大学附属曙光医院四明青年基金项目(SGKJ-201704).

摘　　要：目的探讨丹参酮成分对胆汁酸对小鼠肝微粒体羟化代谢的抑制作用。方法胆汁酸与NADPH在小鼠肝微粒体中共同孵育后,采用液相色谱-串联质谱法(LC-MS/MS)同时检测底物及羟化代谢产物,通过与标准品比对,确定代谢物类型,并将代谢明显的胆汁酸纳入抑制研究。丹参酮ⅡA、隐丹参酮、丹参酮Ⅰ和二氢丹参酮Ⅰ分别与胆汁酸在小鼠肝微粒体中孵育后,同时测定底物及羟化代谢产物浓度,计算IC50值表示对胆汁酸羟化代谢的抑制程度。结果①在小鼠肝微粒体体外孵育体系中,胆汁酸CDCA、LCA、TCDCA、TLCA和TDCA代谢明显,内生清除率分别为0.06、0.11、0.58、1.42和0.26 mL/(min·mg)。②CDCA、LCA、TCDCA和TDCA分别羟化生成α-MCA、MDCA、T-α-MCA和TCA,TLCA羟化生成TDCA、T-α-MCA和T-β-MCA。③二氢丹参酮Ⅰ抑制LCA代谢及代谢物MDCA生成的IC50值分别为0.85和1.04μmol/L,抑制TCDCA代谢及T-α-MCA生成的IC50值分别为1.29和2.74μmol/L,抑制TLCA代谢及TCDCA生成的IC50值分别为0.36和0.66μmol/L;CTS抑制TDCA代谢的IC50值为43.18μmol/L,TSI抑制TCDCA代谢的IC50值为36.19μmol/L。结论丹参酮对胆汁酸在小鼠肝微粒体中的羟化代谢有抑制,其中二氢丹参酮Ⅰ起主要作用,临床使用含有二氢丹参酮Ⅰ制剂时应关注其引起胆汁淤积症的风险。Objective To investigate the inhibition effect of tanshinones on the hydroxylation of bile acids in mouse liver microsomes.Methods The substrates and metabolites were simultaneously measured by LC-MS/MS after bile acids co-incubated with NADPH in mouse liver microsomes.Metabolites were identified by comparison with those of authentic standards,and the bile acids with obvious metabolism were included in the following study.TanshinoneⅡA,crytotanshinone,tanshinoneⅠand dihydrotanshinoneⅠwere respectively incubated with mouse liver microsomes in the presence of bile acids,and the substrates and metabolites were simultaneously measured by LC-MS/MS.The inhibitory effects on hydroxylation of bile acids were evaluated with IC50 values.Results①Bile acids CDCA,LCA,TCDCA,TLCA and TDCA metabolized in mouse liver microsomes,with CLint of 0.06,0.11,0.58,1.42 and 0.26 mL/(min·mg),respectively.②Incubation of CDCA,LCA,TCDCA and TDCA with liver microsomes from C57 BL/6 mice yielded hydroxylated metabolites that were identified asα-MCA,MDCA,T-α-MCA and TCA,respectively,by comparison with authentic standards.TDCA,T-α-MCA and T-β-MCA were detected as hydroxylated metabolites of TLCA.③The IC50 values of dihydrotanshinoneⅠtowards LCA depletion and MDCA generation were 0.85 and 1.04μmol/L,respectively.DihydrotanshinoneⅠinhibited TCDCA depletion and T-α-MCA generation with IC50 values of 1.29 and 2.74μmol/L,respectively.Furthermore,TLCA hydroxylation was inhibited by dihydrotanshinoneⅠwith IC50 values of 0.36μmol/L towards substrate depletion and 0.66μmol/L towards metabolite generation.The IC50 valueof crytotanshinone on TDCA metabolism was 43.18μmol/L.The IC50 value of tanshinoneⅠon TCDCA metabolism was 36.19μmol/L.Conclusion Tanshinones inhibit bile acids hydroxylation in mice liver microsomes.Among them,dihydrotanshinone I plays an important role.Attention should be paid to the potential cholestatic risk of preparations containing dihydrotanshinoneⅠin clinic.

关 键 词：丹参酮 胆汁酸 羟化代谢 肝微粒体 

分 类 号：R285.5[医药卫生—中药学]