摘要
该研究利用Caco-2细胞和正常大鼠考察大麻二酚(cannabidiol,CBD)体外吸收特性和体内药代动力学特征,并进一步研究其抗炎机制。通过CCK-8法确定CBD安全浓度范围,研究时间、浓度、温度、内吞抑制剂和转运抑制剂对CBD跨膜吸收和转运的影响;检测大鼠灌胃给药后不同时间的血药浓度,绘制药-时曲线,计算药代动力学参数;脂多糖(LPS)诱导Caco-2细胞建立炎症损伤模型,检测CBD干预处理后乳酸脱氢酶(LDH)活力、跨膜电阻(TEER)值以及细胞炎症因子水平探究CBD抗炎作用机制。结果表明在实验浓度范围内,Caco-2细胞对CBD的摄取在2 h达到饱和,CBD摄取量和浓度、温度呈正相关,可被内吞进入细胞。CBD可以通过多药耐药蛋白2(MRP2)和乳腺癌耐药蛋白(BCRP)参与的主动转运途径穿透Caco-2细胞,而加入P-gp抑制剂对CBD的转运没有影响。大鼠对CBD吸收迅速,达峰时间tmax为(1.00±0.11)h,在体内的消除较快,t1/2仅为(1.86±0.16)h。另外,CBD显著改善了炎症反应带来的LDH活力升高和TEER降低的现象,可通过降低促炎细胞因子白细胞介素-8(IL-8)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)的表达维持肠道屏障,起到抗炎作用。
This study used Caco-2 cells and normal rats to investigate the in vitro absorption characteristics and in vivo pharmacokinetic characteristics of cannabidiol(CBD)and explore the anti-inflammatory mechanism of CBD.The safe concentration range of CBD was determined by the CCK-8 assay,and then the effects of time,concentration,temperature,endocytosis inhibitors,and transport inhibitors on the transepithelial absorption and transport of CBD were assessed.The blood drug concentration was measured at different time points after oral administration in rats for pharmacokinetic profiling,and the pharmacokinetic parameters were calculated.The Caco-2 cell model of inflammation injury was established with lipopolysaccharide(LPS).The effects of CBD on lactate dehydrogenase(LDH)activity,transendothelial electrical resistance(TEER),and levels of inflammatory cytokines of the modeled cells were examined,on the basis of which the anti-inflammatory mechanism of CBD was deciphered.The results showed that within the concentration range tested in this study,the CBD uptake by Caco-2 cells reached saturation at the time point of 2 h.Moreover,the CBD uptake was positively correlated with concentration and temperature and CBD could be endocytosed into the cells.CBD could penetrate Caco-2 cells through active transport pathways involving multidrug resistance-associate protein 2(MRP2)and breast cancer resistance protein(BCRP),while the addition of P-gp inhibitors had no effect on CBD transport.Rats exhibited rapid absorption of CBD,with the peak time(tmax)of(1.00±0.11)h,and fast elimination of CBD,with a half-life(t1/2)of only(1.86±0.16)h.In addition,CBD significantly ameliorated the increased LDH activity and decreased TEER that were caused by inflammatory response.It maintained the intes-tinal barrier by down-regulating the expression of pro-inflammatory cytokines interleukin-8(IL-8),interleukin-1 beta(IL-1β)and tumor necrosis factor-α(TNF-α),thus exerting anti-inflammatory effects.
作者
李睿
郝瑞
陈珏
路丽艳
李敏
阮文辉
LI Rui;HAO Rui;CHEN Jue;LU Li-yan;LI Min;RUAN Wen-hui(College of Traditional Chinese Medicine and Food Engineering,Shanxi University of Chinese Medicine,Jinzhong 030619,China;Shanxi Center of Drug Evaluation(Shanxi Institute of Medicine and Life Science),Taiyuan 030006,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2024年第17期4777-4785,共9页
China Journal of Chinese Materia Medica
基金
山西省应用基础研究计划项目(201801D221278)
吕梁市引进高层次科技人才重点研发项目(社会发展)(2022RC06)
山西省中医药管理局科研项目(2020ZYYC098)
企业横向项目(2022-05)
山西省科技创新人才团队专项计划(202204051002028)。
