西安交通大学学报（医学版）2024，Vol.45Issue(4)：590-596,7.DOI:10.7652/jdyxb202404010
脱氧胆酸通过ROS/NF-κB通路对Barrett食管细胞氧化应激的影响
Effects of deoxycholic acid on oxidative stress in Barrett's oesophagus cells via ROS/NF-κB pathway
摘要
Abstract
Objective To explore the effects and mechanisms of deoxycholic acid(DCA)on oxidative stress in human BAR-T cells.Methods The human Barrett epithelial cell line BAR-T was cultured in vitro,and the BAR-T cells were intervened with different concentrations of DCA(100,200,and 300 μmol/L)and different action time(30 min,60 min,3 h and 6 h).The mRNA and protein expression of COX-2 were detected by Real-time PCR and Western blotting assay.The intracellular reactive oxygen species(ROS)content was detected by direct microscope observation and flow cytometry,and was compared with that of 200 μmol/L DCA+5 mmol/L NAC(ROS scavenger N-acetylcysteine)group;cellular immunofluorescence was used to detect the entry of cellular p65 protein into the nucleus,which was then compared with 200 μmol/L DCA+PDTC(100 μmol/L,NF-κB pathway inhibitor)group.Results Compared to the control group,DCA increased ROS content in BAR-T cells in a dose-dependent manner.Additionally,5 mmol/L NAC significantly inhibited DCA-induced ROS release.Furthermore,both 200 μmol/L and 300 μmol/L DCA significantly increased COX-2 mRNA expression compared to the control group at the same intervention time.The 6 h groups for both 200 μmol/L and 300 μmol/L DCA showed a significant increase in COX-2 mRNA expression compared to the 1 h group.Compared to the control group,treatment with 200 μmol/L DCA significantly increased the expression of COX-2 protein.Meanwhile,200 μmol/L DCA promoted the nucleation of p65 protein,which was inhibited by PDTC.Conclusion DCA activates the NF-κB signaling pathway and upregulates the expression of COX-2 by elevating the intracellular ROS expression and promoting the entry of p65 protein into the nucleus.关键词
脱氧胆酸(DCA)/Barrett细胞/氧化应激/NF-κB信号通路Key words
deoxycholic acid(DCA)/Barrett cell/oxidative stress/NF-κB signaling pathway分类
医药卫生引用本文复制引用
冯诚,吕建瑞,王瑾,张军..脱氧胆酸通过ROS/NF-κB通路对Barrett食管细胞氧化应激的影响[J].西安交通大学学报（医学版）,2024,45(4):590-596,7.基金项目
国家卫生部临床重点项目(No.2007353) （No.2007353）
陕西省自然科学基金青年项目(No.2020JQ-557)Supported by the Clinical Key Project of the Ministry of Health(No.2007353)and Shaanxi Provincial Natural Science Foundation Youth Project(No.2020JQ-557) （No.2020JQ-557）
