染料木素-川芎嗪共晶在Caco-2细胞模型中的转运研究  

Study on Transport of Genistein-Tetramethylpyrazine Cocrystal in Caco-2 Cell Model

作　　者：张雨晴 郝星慧 王飞[1] 刘梦歌 赵兴华[1] 何欣[1] ZHANG Yu-qing;HAO Xing-hui;WANG Fei;LIU Meng-ge;ZHAO Xing-hua;HE Xin(College of Veterinary Medicine,Hebei Agricultural University,Baoding 071000,China)

机构地区：[1]河北农业大学动物医学院,河北保定071000

出　　处：《动物医学进展》2024年第2期40-47,共8页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(32002326)。

摘　　要：旨在考察染料木素(genistein,GEN)、川芎嗪(tetramethylpyrazine,TMP)、染料木素-川芎嗪共晶(GEN-TMP)及染料木素与川芎嗪的物理混合物(GEN+TMP)在Caco-2细胞模型中的转运特征;建立Caco-2细胞模型,并以细胞跨膜电阻和标志物渗漏检查等指标进行验证,采用高效液相色谱法,考察并计算安全浓度下药物的累积转运量、表观渗透系数和外排率,并探讨P糖蛋白(P-gp)抑制剂维拉帕米、乳腺癌耐药蛋白(breast cancer resistant protein,BCRP)抑制剂KO143和多药耐药相关蛋白2(multidrug resistance-associated protein 2,MRP2)抑制剂MK571对转运的影响。结果显示,Caco-2细胞模型完整性与功能性良好,GEN浓度为40μg/mL时,GEN、TMP、GEN-TMP以及GEN+TMP的细胞存活率分别为90.06%、84.21%、97.60%和89.37%;GEN、TMP、GEN-TMP和GEN+TMP的表观渗透系数(P_(app))大于1.0×10^(-6)cm/s,属于吸收良好药物;GEN-TMP中GEN的累积转运量和P_(app)值分别为(2.78±0.11)μg和(8.61±0.33)×10^(-6)cm/s,比GEN的(1.92±0.15)μg和(5.96±0.47)×10^(-6)cm/s提高了44.79%和44.46%;GEN+TMP中GEN的累积转运量和P_(app)值与GEN无显著性差异。GEN只受BCRP的外排作用,GEN-TMP中的GEN同时受到P-gp和BCRP的外排作用,TMP、GEN-TMP和GEN+TMP中的TMP均受到MRP2的外排作用。结果表明,相同浓度下,GEN-TMP的细胞存活率高于GEN+TMP。GEN-TMP的吸收强于GEN和GEN+TMP中的GEN,共晶受外排蛋白的作用区别于GEN和GEN+TMP中的GEN,研究工作为共晶的转运研究提供了借鉴和参考。The aim was to investigate the transport characteristic of genistein(GEN),tetramethylpyrazine(TMP),the cocrystal GEN-TMP and the physical mixture GEN+TMP in Caco-2 cells model.The integrity of the cell monolayer was verified by measuring the transepithelial electrical resistance(TEER)and marker penetrant inspection.Concentrations of samples were determined by high-performance liquid chromatography(HPLC).The cumulative transport capacity,apparent permeability coefficient(P_(app))and efflux rate of drugs were calculated.The efflux inhibitors such as P-glycoprotein(P-gp)inhibitor Verapamil,breast cancer resistance protein(BCRP)inhibitor KO143,and multidrug resistance protein 2(MRP2)inhibitor MK571 were added to explore the effects to transport mechanism.The results showed that the cell model of Caco-2 has good integrity and functionality.When the GEN concentration was 40μg/mL,the cell viability of GEN,TMP,cocrystal GEN-TMP and physical mixture GEN+TMP were 90.06%,84.21%,97.60%and 89.37%respectively,the P_(app)of those were all higher than 1.0×10^(-6) cm/s,which indicated drugs with good absorption.The cumulative transport and P_(app)of GEN were(1.92±0.15)μg and(5.96±0.47)×10^(-6)cm/s,and GEN in GEN-TMP were(2.78±0.11)μg and(8.61±0.33)×10^(-6)cm/s.Compared with GEN,GEN form GEN-TMP were increased 44.79%and 44.46%respectively.But,there was no significant difference between GEN form GEN+TMP and GEN.GEN was affected by the efflux of BCRP and GEN in GEN-TMP was affected by the efflux of P-gp and BCRP.The transport of TMP,TMP in GEN-TMP and TMP in GEN+TMP were all influenced by MRP2.These results indicated that the cell viability of GEN-TMP was higher than that of GEN+TMP at the same concentration,the absorption of GEN-TMP was better than that in GEN and GEN+TMP and the effects of efflux inhibitors of GEN in GEN-TMP was drastically different from GEN and GEN in GEN+TMP.This study provided a reference for the transport study of cocrystal.

关 键 词：CACO-2细胞模型 染料木素 川芎嗪 共晶 转运机制 

分 类 号：S853.7[农业科学—临床兽医学]