基于LPS诱导的体外炎症模型建立羊耳菊抗炎活性成分的PK-PD结合模型  

Establishment of PK-PD model in anti-inflammatory active components in Inula cappa extract based on lipopolysaccharide-induced in vitro inflammation model

作　　者：周杰 张青 陈艺 薛寸 李月婷 黄勇 郑林 黄静 陈思颖 巩仔鹏 ZHOU Jie;ZHANG Qing;CHEN Yi;XUE Cun;LI Yue-ting;HUANG Yong;ZHENG Lin;HUANG Jing;CHEN Si-ying;GONG Zi-peng(Provincial Key Laboratory of Pharmaceutics in Guizhou Province,State Key Laboratory of Functions and Applications of Medicinal Plants,Engineering Research Center for the Development and Application of Ethnic Medicine and Traditional Chinese Medicine(Ministry of Education),School of Pharmacy,Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]贵州医科大学药学院、贵州省药物制剂重点实验室、省部共建药用植物功效与利用国家重点实验室、民族药与中药开发应用教育部工程研究中心,贵州贵阳550004

出　　处：《中国中药杂志》2022年第23期6308-6319,共12页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81860734);贵州省大学生创新创业计划项目(S202110660046);国家大学生创新创业计划项目(20195200132);贵州医科大学优秀青年人才计划项目(2022-104)。

摘　　要：该文旨在基于脂多糖(LPS)诱导的体外炎症模型建立羊耳菊抗炎活性成分的PK-PD结合模型,以期阐明羊耳菊抗炎活性成分在炎症细胞内的动态变化与其药效消长之间的相互关系。首先采用1μg·mL^(-1)LPS诱导RAW264.7细胞24 h后,建立体外炎症模型。然后比较400μg·mL^(-1)的羊耳菊提取物干预后其含有的木犀草苷(luteolin,LUT)、绿原酸(chlorogenic acid,CA)、隐绿原酸(cryptochlorogenic acid,CCA)、3,4-二咖啡酰基奎宁酸(3,4-dicaffeoylquinic acid,3,4-DCQA)、4,5-二咖啡酰基奎宁酸(4,5-dicaffeoylquinic acid,4,5-DCQA)等5个抗炎活性成分在正常细胞和炎症细胞中的药代动力学。同时取各时间点细胞上清液测定炎症因子NO和TNF-α作为药效研究,与药动学部分通过WinNonlin 8.2软件中的Phoenix Model进行拟合,建立羊耳菊提取物中LUT、CA、CCA、3,4-DCQA、4,5-DCQA等5个成分的整合PK-PD结合模型。结果表明,与正常细胞相比,5个成分在模型细胞中摄取增加或下降,T_(max)提前,吸收变快,MRT、t_(1/2)延长,CL_(z/F)和V_(z/F)有增加或减少的趋势。以NO为药效指标时,羊耳菊中多效应成分整合后的PK-PD模型为E=7.45×[1-Ce^(5.74)/(78.245.74+Ce^(5.74))];以TNF-α为药效指标时,羊耳菊中多效应成分整合后的PK-PD模型为E=79.28×[1-Ce^(6.45)/(85.106.45+Ce^(6.45))]。研究结果提示炎症状态会改变羊耳菊的细胞药代动力学过程,而且羊耳菊活性成分发挥抗炎的作用可能与下调炎症细胞中NO和TNF-α的分泌有关,且NO和TNF-α可能为羊耳菊活性部位作用于炎症相关作用靶点。In the present study,a pharmacokinetics(PK)-pharmacodynamics(PD)model in the anti-inflammatory active components in Inula cappa extract was established based on the lipopolysaccharide(LPS)-induced in vitro inflammation model in order to clarify the relationship between the dynamic changes of anti-inflammatory active components in inflammatory cells and their efficacy.Firstly,the inflammation model in vitro was induced by 1μg·mL^(-1)LPS in RAW264.7 cells for 24 h.After treatment with 400μg·mL^(-1)I.cappa extract,the pharmacokinetics(PK)of five anti-inflammatory active components,including luteolin(LUT),chlorogenic acid(CA),cryptochlorogenic acid(CCA),3,4-dicaffeoylquinic acid(3,4-DCQA),and 4,5-dicaffeoylquinic acid(4,5-DCQA),in normal cells and inflammatory cells was compared.Meanwhile,the PD study was carried out by measuring the inflammatory factors NO and TNF-αin the cell supernatant at each time point,which was fitted with PK by the Phoenix Model in the WinNonlin 8.2 to establish the PK-PD model for five components including LUT,CA,CCA,3,4-DCQA,and 4,5-DCQA.The results showed that compared with normal cells,the model cells showed increased or decreased uptake of five components,advanced T_(max),faster absorption,prolonged MRT and t_(1/2),and increasing or decreasing trend of CL_(z/F)and V_(z/F).When NO was used as the efficacy index,the PK-PD model after the integration of the multi-effect components in I.cappa was E=7.45×[1-Ce^(5.74)/(78.245.74+Ce^(5.74))],while with TNF-αas the efficacy index,the PK-PD model after the integration of the multi-effect components in I.cappa was E=79.28×[1-Ce^(6.45)/(85.106.45+Ce^(6.45))].The results of the study suggested that the inflammatory state could change the cellular PK of I.cappa.The anti-inflammatory effect of active components in I.cappa might be related to the down-regulation of the secretion of NO and TNF-αin inflammatory cells,and NO and TNF-αmight serve as the anti-inflammatory targets of active components of I.cappa.

关 键 词：羊耳菊 抗炎活性成分 炎症细胞模型 细胞药代动力学 PK-PD模型 

分 类 号：R285[医药卫生—中药学]