中国临床药学杂志2025，Vol.34Issue(5)：329-335,7.DOI:10.19577/j.1007-4406.2025.05.002
药物血-前列腺屏障细胞模型构建及occludin蛋白的作用研究
Drug blood-prostate barrier cell model construction and investigation of the role of occludin protein
摘要
Abstract
AIM To establish stable root wolff prostate epithelial-1(RWPE-1)and whitmore-prostate mesenchymal Y-1(WPMY-1)overexpressing occludin,investigating the impact of occludin on prostate barrier function and its potential regulatory mechanisms.METHODS The coding sequence of occludin was cloned into a lentiviral vector,and viral particles were produced in 293T cells to infect RWPE-1 and WPMY-1 cells.Stable transfectants were selected using puromycin.Infection efficiency and protein expression in the occludin-related pathway were assessed using quantitative PCR(qPCR)and Western blot.Prostate cell models of drugs were constructed using Transwell inserts to investigate the barrier function.The cell models were divided into control group,WPMY-1 group,RWPE-1 group,CO-R/W group,WPMY-1 with occludin overexpression(ov-WPMY)group,RWPE-1 with occludin overexpression(ov-RWPE)group,ov-CO-R/W group.Barrier integrity was evaluated by measuring transendothelial electrical resistance and performing fluorescent dye transport assays.Evaluation of the drug transport properties of cefradine and levofloxacin was conducted under different conditions.RESULTS Stable cell lines overexpressing occludin(ov-RWPE and ov-WPMY)were successfully established.Levels of c-Jun N-terminal kinase 2(JNK2)and apoptosis signal-regulating kinase 1(ASK1)in ov-RWPE and ov-WPMY groups were significantly higher than those in the RWPE-1 group and WPMY-1 group(P＜0.01).Prostate barrier models were successfully constructed.Compared to CO-R/W group,the drug permeability of cefradine and levofloxacin in the ov-CO-R/W group was significantly lower(P＜0.01).CONCLUSION Prostate stromal cells are involved in constituting the blood-prostate barrier,and occludin may be a key protein in constituting the barrier.关键词
血-前列腺屏障/药物分布/occludin蛋白Key words
blood-prostate barrier/drug distribution/occludin分类
药学引用本文复制引用
陈缘静,葛蕊,韩秀娟,朱伟强,杨云云,王卓..药物血-前列腺屏障细胞模型构建及occludin蛋白的作用研究[J].中国临床药学杂志,2025,34(5):329-335,7.基金项目
国家自然科学基金(编号81870520) （编号81870520）
