肝毒性剂量的吴茱萸入血成分浓度-毒性分析

Concentration-toxicity analysis of hepatotoxicity induced by Evodia rutaecarpa

关键词:  吴茱萸  柠檬苦素  肝毒性  血药浓度

Key words:  Evodia  Rutaecarpa, limonin, hepatotoxicity, plasma  concentration

摘要:

目的 基于吴茱萸致肝毒性,分析体内血药浓度-毒性关系,进一步揭示吴茱萸的肝毒性。方法 采用UPLC测定吴茱萸水提物中吴茱萸碱、吴茱萸次碱和柠檬苦素的含量；采用LC-MS/MS测定吴茱萸水提物和柠檬苦素致小鼠肝毒性动物血浆中柠檬苦素的浓度,动物分为吴茱萸水提物灌胃组、柠檬苦素灌胃组、柠檬苦素腹腔注射组,给药剂量分别为60 g.kg_-1^{,200 mg}.kg_-1 ^{和 200 mg}.kg_-1^,各组于给药前和给药后不同时间点采血检测血浆中柠檬苦素的浓度。结果 吴茱萸水提物中吴茱萸碱、吴茱萸次碱、柠檬苦素含量分别为0.08%、0.04%、1.09%。吴茱萸水提物和柠檬苦素致小鼠肝毒性血药浓度检测显示,柠檬苦素灌胃组在未引起明显肝毒性情况下血浆柠檬苦素浓度和体内暴露量最低,C_max为32.1±14.5 ng.mL_-1^、AUC_(0-t)为138.5±139.3 h.ng.mL_-1^,同等剂量的柠檬苦素腹腔注射组在引起明显肝毒性情况下血浆柠檬苦素浓度和体内暴露量均增加,C_max为1080.0±296.3 ng.mL_-1^、AUC_(0-t)为10234.5±6917.3 h.ng.mL_-1^,吴茱萸水提物灌胃组在引起明显肝毒性情况下血药浓度和体内暴露量最高,C_max为11130.0±3047.8 ng.mL_-1^、AUC_(0-t)为91797.2±31524.4 h.ng.mL_-1^。结论 吴茱萸引起的肝毒性越明显,其血浆柠檬苦素的浓度越高,柠檬苦素是吴茱萸质量控制的关键标志物。

Abstract:

OBJECTIVE To analyse the plasma concentration-toxicity relationship in vivo based on the hepatotoxicity of Evodia rutaecarpa and to further reveal the hepatotoxicity. METHODS The contents of evodiamine, rutaecarpine and limonin in the aqueous extract of Evodia rutaecarpa were determined using ultra high performance liquid chromatography (UPLC). The plasma concentration of limonin was determined in mice with hepatotoxicity induced by Evodia rutaecarpa aqueous extract and limonin using liquid chromatography tandem mass spectrometry (LC-MS/MS). The mice were divided into three groups: the Evodia rutaecarpa aqueous extract group, which was given a gavage dose of 60 g?kg-1, and two limonin groups, one given a gavage dose of 200 mg?kg-1 and the other an intraperitoneal injection at the same dose. The plasma concentration of limonin was measured at various time points before and after administration via blood collection. RESULTS The contents of evodiamine, rutaecarpine and limonin in the aqueous extract of Evodia rutaecarpa were 0.08%, 0.04% and 1.09%, respectively. Plasma concentration tests showed that the gavage group of limonin had the lowest Cmax and AUC(0-t) without causing significant hepatotoxicity, with a Cmax of 32.1±14.5 ng?mL-1 and an AUC(0-t) of 138.5±139.3 h?ng?mL-1, and the same dose of limonin injected intraperitoneally group increased both Cmax and AUC(0-t) in case of causing significant hepatotoxicity, with a Cmax of 1080.0±296.3 ng?mL-1 and an AUC(0-t) of 10234.5±6917.3 h?ng?mL-1, and the highest Cmax and AUC(0-t) were observed in the gavage group of the aqueous extract of Evodia rutaecarpa in the case of causing significant hepatotoxicity, with a Cmax of 11130.0±3047.8 ng?mL-1 and an AUC(0-t) of 91797.2±31524.4 h?ng?mL-1. CONCLUSION The more severe the hepatotoxicity caused by Evodia rutaecarpa, the higher limonin plasma concentration, and limonin is a key marker for quality control of Evodia rutaecarpa.