摘要
目的:基于脾胃虚寒大鼠,鉴定厚朴温中汤体内外化学成分,并分析指标性成分的药代动力学性质。方法:采用超高效液相色谱-四极杆-飞行时间串联质谱法(UPLC-Q-TOF-MS/MS)对厚朴温中汤中的化学成分进行分析与鉴定。从18只SD大鼠中随机选取6只作为空白组,其余大鼠采用猪油和冷食醋长时间灌胃构建脾胃虚寒型大鼠模型,造模成功后随机平均分为模型组和厚朴温中汤组(13.5 g·kg^(-1),以生药计),给药组灌胃给予相应剂量厚朴温中汤药液,空白组与模型组灌胃等体积蒸馏水。采用酶联免疫吸附测定法(ELISA)检测各组大鼠血清胃泌素(GAS)和胃动素(MTL)含量。同时,于给药后不同时间点收集血浆样品,采用UPLC-Q-TOF-MS/MS分析厚朴温中汤的入血原型成分与代谢产物,并结合超高效液相色谱-三重四极杆/线性离子阱质谱法(UPLC-QTRAP-MS/MS)检测厚朴温中汤中4个指标性成分厚朴酚、和厚朴酚、山姜素及橙皮苷的血药浓度,利用DAS 2.0软件计算药代动力学参数。结果:厚朴温中汤中共鉴定出化学成分79个,包括黄酮类化合物44个,以及木脂素类化合物11个。鉴定到入血原型成分18个,代谢物27个,主要代谢途径为葡萄糖醛酸化、硫酸化、氧化及水解等,主要代谢类型为Ⅰ相及Ⅱ相代谢。药代动力学结果显示,4个指标性成分中厚朴酚及和厚朴酚的达峰时间(t_(max))一致,表现出相似的药物代谢特征;山姜素的t_(max)最短,吸收速率最快,最早达到血药浓度峰值水平;橙皮苷在大鼠体内平均驻留时间(MRT_(0-t))最短且代谢速率最快。结论:本研究明确了厚朴温中汤在脾胃虚寒大鼠模型中的入血原型及其代谢产物,揭示了主要活性成分的药代动力学特征,可为厚朴温中汤的药效物质基础研究与治疗脾胃虚寒证的临床应用提供科学依据。
Objective:To identify the chemical components of Houpo Wenzhongtang in vivo and in vitro and to analyze the pharmacokinetic properties of the index components in rats with deficiency-cold of spleen and stomach.Method:The chemical components of Houpo Wenzhongtang was analyzed and identified by ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-QTOF-MS/MS).Six rats were randomly selected from 18 SD rats as the blank group,and the remaining rats were given lard and cold vinegar for a long time to construct a rat model with deficiency-cold of spleen and stomach.After successful modeling,the rats were randomly divided into the model group and Houpu Wenzhongtang group(13.5 g·kg^(-1),calculated as crude drug).The administration group was given the corresponding dose of Houpu Wenzhongtang by gavage,and the blank group and the model group were given the same amount of distilled water by gavage.Enzyme-linked immunosorbent assay(ELISA)were used to measure gastrin(GAS)and motilin(MTL)levels in each group.At the same time,plasma samples were collected at different time points after administration,and blood-entry prototype components and metabolites of Houpo Wenzhongtang were analyzed by UPLC-Q-TOF-MS/MS.On this basis,plasma concentrations of magnolol,honokiol,alpinetin and hesperidin in Houpo Wenzhongtang were determined by ultra performance liquid chromatography coupled with triple quadrupole/linear ion trap mass spectrometry(UPLC-QTRAP-MS/MS),and the pharmacokinetic parameters were calculated using DAS 2.0 software.Result:A total of 79 chemical components,including 44 flavonoids and 11 lignans,were identified in Houpo Wenzhongtang.Meanwhile,18 blood-entry prototype components and 27 metabolites were identified,the main metabolic pathways of metabolites were glucuronidation,sulfation,oxidation and hydrolysis,and phaseⅠand phaseⅡwere the two primary forms of metabolism.Pharmacokinetic results showed that among the four index components,the time to peak(t_(max))values of magnolol and honokiol were consistent and exhibited similar drug metabolism characteristics,the t_(max) of alpinetin was the shortest,and the absorption rate was the fastest,which had the earliest peak plasma concentration levels,and hesperidin had the shortest mean residence time(MRT(0-t))and the highest metabolic rate in rats.Conclusion:This study clarifies the blood-entry prototype components and their metabolites of Houpo Wenzhongtang in the rat model of deficiency-cold of spleen and stomach,and reveals the pharmacokinetic characteristics of the main active ingredients,which can provide a scientific basis for the study of pharmacodynamic material basis of this formula and its clinical application in treating the syndrome of deficiency-cold of spleen and stomach.
作者
石宣宜
陈家毅
陈殊
顾子娴
欧阳莉
罗光燕
彭晓顺
刘建群
SHI Xuanyi;CHEN Jiayi;CHEN Shu;GU Zixian;OUYANG Li;LUO Guangyan;PENG Xiaoshun;LIU Jianqun(Key Laboratory of Modern Preparation of Traditional Chinese Medicine of Ministry of Education,Jiangxi University of Chinese Medicine,Nanchang 330004,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2024年第17期145-154,共10页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(82160731)
江西中医药大学校级科技创新团队发展计划项目(CXTD22007)。
关键词
厚朴温中汤
化学成分
入血成分
代谢产物
药代动力学
液质联用技术
脾胃虚寒
Houpo Wenzhongtang
chemical components
blood-entry components
metabolites
pharmacokinetics
liquid chromatography-mass spectrometry
deficiency-cold of spleen and stomach
