洋金花山柰酚-7-O-吡喃鼠李糖苷在Caco-2细胞模型中的吸收转运机制研究  

Mechanism Study on Uptake and Transportation of Kaempferol 7-O-α-L-rhamnopyranosid in Caco-2 cells

作　　者：张慧[1] 米晓 孟健男[1] 李毅翔 匡海学 王艳艳[1] ZHANG Hui;MI Xiao;MENG Jian-nan;LI Yi-xiang;KUANG Hai-xue;WANG Yan-yan(Heilongjiang University of Chinese Medicine,Harbin 150040,China;Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences,Beijing 100193,China;Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]黑龙江中医药大学,黑龙江哈尔滨150040 [2]中国医学科学院药用植物研究所,北京100193 [3]广东药科大学,广东广州510006

出　　处：《时珍国医国药》2022年第1期60-63,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(82003974);黑龙江省青年科学基金项目(QC2013C085,LH2019H106);黑龙江省教育厅科学技术研究项目(12531623);黑龙江省哈尔滨市科技局项目(2013RFQXJ070)。

摘　　要：目的阐明洋金花黄酮类化合物山柰酚-7-O-α-L-吡喃鼠李糖苷(Kaempferol 7-O-α-L-rhamnopyranoside,K1)吸收转运机制。方法首先,探究K1浓度对IFN-γ所致HaCaT细胞损伤的影响,之后应用UPLC-PDA法对洋金花中黄酮类化合物K1在人源肠Caco-2细胞单层模型中的吸收转运特性进行定量分析,分别考察pH、浓度、时间节点等因素对化合物转运的影响,并在最佳条件下加入P-糖蛋白(P-glycoprotein,P-gp)抑制剂维拉帕米和MRP抑制剂MK-571考察两者在K1转运中的作用。结果不同浓度K1能显著增加IFN-γ诱导的HaCaT细胞活性(P<0.01),在pH 7.4的环境值下表观渗透系数(Papp)达到最大,转运时长为2 h时,药物累计转运量最大。K1的Papp(BL→AP)随浓度的增加而增加,外排率(ER)均处于0.5-1.5,显示为被动转运,加入P-gp和MRP抑制剂后,ER值降低。结论洋金花黄酮类化合物K1在Caco-2细胞中以被动扩散为主,药物扩散过程中有P-gp糖蛋白和MRP参与。Objective To study the mechanism on uptake and transportation of Kaempferol 7-O-α-L-rhamnopyranosid(K1)in Caco-2 cells.Methods First,we explored the concentration of K1 how to effect on HaCaT cells induced by IFN-γ.Then,UPLC-PDA method was used to quantitatively analyze transport characteristics of K1 in Caco-2 cell model,and transportation experiment was conducted.Then,the effects of pH,concentration,and time on the transportation of K1 were investigated.Under the optimal conditions,the effects of P-glycoprotein(P-gp)inhibitor verapamil and MRP inhibitor MK-571 on the uptake and transportation of K1 in the sample were studied in Caco-2 cells.Results Different concentrations of K1 could significantly increase the activity of HaCaT cells induced by IFN-γ and the apparent permeability coefficient(Papp)of K1 reached the maximum at pH 7.4.When the transport time was 2 h,the cumulative drug transport volume was the largest.The Papp(BL→AP)of K1 increased with the increase of concentration,and the effluent rate(ER)was between 0.5 and 1.5,indicating passive diffusion.However,after the addition of p-gp and MRP inhibitors,the ER decreased.Conclusion K1,a compound of Datura metel L.,was mainly passively diffused in Caco-2 cells,and P-gp and MRP were involved in diffusion process.

关 键 词：洋金花 山柰酚苷 CACO-2细胞 

分 类 号：R285.5[医药卫生—中药学]