莱柯霉素在Caco-2细胞单层模型中的转运特性研究

doi:10.16431/j.cnki.1671-7236.2023.12.031

Abstract

Abstract: 【Objective】 This experiment was aimed to investigate the transport characteristics of lekethromycin across Caco-2 cell monolayer model,and to provide further experimental evidence for further understanding of the absorption process of lekethromycin in vivo.【Method】 A Caco-2 cell monolayer model was established to verify the compactness of the Caco-2 cell monolayer model and to screen the model by measuring the transmembrane resistance (TEER) value and the apparent permeability Papp_(AP-BL) of the low permeability drug fluorescein sodium permeability test.The transport characteristics of lekethromycin from the apical side (AP) to the basolateral side (BL) and from the basolateral side to the apical side in the Caco-2 cell monolayer model were investigated,and the effects of three factors,drug concentration,pH and efflux transporter (P-gp,MRP2 and BCRP) inhibitors on the transportation of lekethromycin in Caco-2 monolayer model were analyzed.The drug concentrations were measured by UPLC-MS/MS,and its apparent permeability and efflux ratio were calculated.【Result】 The resistance value test and sodium fluorescein leakage test for 20 days showed that with the increase of culture time,the transmembrane resistance value of Caco-2 cell monolayer model gradually increased and reached about 400 Ω·cm² at the 20th day.The linear relationship of fluorescein sodium was good within 0.1-6 μg/mL (R²=0.991),and the calculated fluorescein sodium Papp_(AP-BL) was (1.57×10^-7±9.92×10^-9) cm/s,which was less than 0.5×10^-6 cm/s specified in the test.The Caco-2 cell monolayer model was successfully established and could be used to study the transport characteristics of lekethromycin.Lekethromycin Papp_(AP-BL) values did not change appreciably over time and were low,while Papp_(BL-AP) values increased over time,reaching a maximum level of 2.0350×10^-6 cm/s at 120 min of incubation,and intestinal efflux of lekethromycin was strong.Papp_(BL-AP) values increased with increasing concentrations of lekethromycin in a concentration-dependent manner.When the pH of the solution was 8.0,the Papp_(BL-AP) value (39.3871×10^-6 cm/s) was significantly higher than that of the solutions at pH 6.5 and 7.4,and alkaline intestinal fluid accelerated lekethromycin efflux.Verapamil,probenecid,and GF120918 inhibitors all significantly inhibited the lateral transport of lekethromycin from BL to AP,with efflux ratios greater than 1.5,and efflux proteins might be involved in transport.【Conclusion】 Lekethromycin showed poor transport and low absorption rate in the in vitro Caco-2 cell monolayer model,showing low permeability compounds.With the increase of concentration and pH,efflux was enhanced,efflux transporters P-gp,MRP2 and BCRP might be involved in the transport process of lekethromycin.

Key words: Caco-2 cell monolayer model; lekethromycin; transport; apparent permeability

CLC Number:

S859.7

WEN Zeyu, XIAO Hongzhi, SUN Pan, LIU Yu, YANG Yuxin, WANG Yue, LI Jing, ZHANG Lu, GONG Xiaohui, ZHOU Degang, CAO Xingyuan. Transport Characteristics of Lekethromycin Across Caco-2 Cell Monolayer Model[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(12): 5094-5103.

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Transport Characteristics of Lekethromycin Across Caco-2 Cell Monolayer Model

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