摘要
该研究旨在探究中医经典名方血府逐瘀汤(Xuefu Zhuyu Decoction, XFZY)与阿托伐他汀(atorvastatin, AT)的中西药相互作用。采用RT-PCR分析LS174T细胞的药物代谢和转运相关蛋白转录水平;测定不同底物浓度和孵育时长下胞内药物摄取量,优化转运体MDR1特异探针罗丹明123及AT的模型反应条件,建立人肠道药物相互作用研究细胞模型;CCK-8法评价XFZY对LS174T细胞毒,在单次及连续48 h XFZY培养后再加入AT或罗丹明123共孵育,通过测定胞内药物浓度及相关转运体和代谢酶转录水平,分析XFZY对AT人肠道吸收的影响及机制。体外实验结果显示,单次高浓度的XFZY共培养,可使罗丹明123和AT胞内浓度均显著上升;48 h高浓度XFZY共培养升高了AT的摄取水平,显著诱导CYP3A4、UGT1A1基因表达水平,抑制了OATP2B1基因表达水平。为与体外人源细胞模型评估结果进行比较,采用大鼠进行XFZY与AT合并用药的药代动力学实验。SD大鼠随机分为空白对照组及XFZY给药组,连续灌胃给药14 d后合并给予AT,LC-MS/MS检测药后血浆样品中AT及代谢产物2-羟基阿托伐他汀酸(2-HAT)、4-羟基阿托伐他汀酸(4-HAT)、阿托伐他汀内酯(ATL)、2-羟基阿托伐他汀内酯(2-HATL)、4-羟基阿托伐他汀内酯(4-HATL)的浓度,计算药代动力学参数。大鼠药代动力学分析表明,连续给药XFZY没有显著改变大鼠体内AT的药代动力学特征,但AT及代谢物2-HAT、4-HAT、2-HATL的AUC0-6 h分别升高了21.37%、14.94%、12.42%、6.68%,主要代谢产物的代谢率有下降的趋势。该研究提示与XFZY联用可显著提高AT在人肠细胞中的摄取水平,不同程度地升高大鼠体内AT及主要代谢产物的暴露水平,其机制可能主要来自对肠道MDR1转运活性的抑制。
The study aims to explore the herb-drug interaction between Xuefu Zhuyu Decoction(XFZY)and atorvastatin(AT).Reverse transcription polymerase chain reaction(RT-PCR)was used to analyze the transcription levels of proteins related to drug metabolism and transport in LS174T cells,detect the intracellular drug uptake under various substrate concentrations and incubation time,and optimize the model reaction conditions of transporter multidrug resistance protein 1(MDR1)-specific probe Rhodamine 123 and AT to establish a cell model for investigating the human intestinal drug interaction.The cell counting kit-8(CCK-8)method was adopted to evaluate the cytotoxicity of XFZY on LS174T cells.After a single and continuous 48 h culture with XFZY,AT or Rhodamine 123 was added for co-incubation.The effect and mechanism of XFZY on human intestinal absorption of AT were analyzed by measuring the intracellular drug concentrations and transcription levels of related transporters and metabolic enzymes.The results of in vitro experiments show that a single co-culture with a high concentration of XFZY significantly increases the intracellular concentrations of Rhodamine 123 and AT.A high concentration of XFZY co-culture for 48 h increases the AT uptake level,significantly induces the CYP3A4 and UGT1A1 gene expression levels,and inhibits the OATP2B1 gene expression level.To compare with the evaluation results of the in vitro human cell model,the pharmacokinetic experiment of XFZY combined with AT was carried out in rats.Sprague-Dawley(SD)rats were randomly divided into a blank control group and an XFZY group.After 14 days of continuous intragastric administration,AT was given in combination.The liquid chromatography-mass spectrometry(LC-MS)/MS method was used to detect the concentrations of AT and metabolites 2-hydroxyatorvastatin acid(2-HAT),4-hydroxyatorvastatin acid(4-HAT),atorvastatin lactone(ATL),2-hydroxyatorvastatin lactone(2-HATL),and 4-hydroxyatorvastatin lactone(4-HATL)in plasma samples,and the pharmacokinetic parameters were calculated.Pharmacokinetic analysis in rats shows that continuous administration of XFZY does not significantly change the pharmacokinetic characteristics of AT in rats,but the AUCo-6h values of AT and metabolites 2-HAT,4-HAT,and 2-HATL increase by 21.37%,14.94%,12.42%,and 6.68%,respectively.The metabolic rate of the main metabolites shows a downward trend.The study indicates that administration combined with XFZY can significantly increase the uptake level of AT in human intestinal cells and increase the exposure level of AT and main metabolites in rats to varying degrees.The mechanism may be mainly due to the inhibition of intestinal MDR1 transport activity.
作者
李想
易欢
任常英
郭好好
杨宏天
张颖
LI Xiang;YI Huan;REN Chang-ying;GUO Hao-hao;YANG Hong-tian;ZHANG Ying(Beijing Key Laboratory of Chinese Medicine Pharmacology,Institute of Basic Medical Sciences,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China)
出处
《中国中药杂志》
北大核心
2025年第11期3159-3167,共9页
China Journal of Chinese Materia Medica
基金
中国中医科学院科技创新工程项目(CI2021A04906)
北京市卓越临床研究计划项目(BRWEP2024Z014170102)
国家自然科学基金项目(81873179)。
