Role and mechanism of COX-2 in glycochenodeoxycholate-induced apoptosis of mouse extrahepatic biliary epithelial cells
10.3760/cma.j.cn431274-20240328-00538
- VernacularTitle:COX-2在甘氨鹅脱氧胆酸盐诱导的小鼠肝外胆管上皮细胞凋亡中的作用及机制
- Author:
Yuxiang ZHOU
1
;
Denghui LIU
1
;
Zhao HUANG
1
;
Qiangxing XIANG
1
;
Xianglian TANG
1
Author Information
1. 中南大学湘雅医学院附属儿童医院（湖南省儿童医院）普外科，长沙 410007
- Publication Type:Journal Article
- Keywords:
Cyclooxygenase 2;
Bile ducts, extrahepatic;
Epithelial cells;
Apoptosis;
Autophagy
- From:
Journal of Chinese Physician
2025;27(5):682-687
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the role of cyclooxygenase-2 (COX-2) in glycochenodeoxycholic acid (GCDC)-induced apoptosis of mouse extrahepatic biliary epithelial cells (EBECs) and clarify its possible mechanism.Methods:EBECs were cultured in vitro and infected with RNAi-COX-2 lentivirus (GCDC+ shCOX-2 group). EBECs were then treated with different concentrations (0, 25, 50, 100, 200, 400, 800 μmol/L) of GCDC (GCDC group). Cell proliferation activity was detected by cell counting kit-8 (CCK-8) assay; lactate dehydrogenase (LDH) release rate was measured by colorimetry; apoptosis was analyzed by flow cytometry; caspase-3 activity was detected by fluorescent probe method; COX-2 mRNA expression was determined by real-time quantitative polymerase chain reaction (qRT-PCR); protein levels of COX-2, autophagy-related proteins LC3, p62, Beclin-1, and apoptosis-related proteins Bax, Bcl-2, cleaved caspase-3 were evaluated by Western blot.Results:Compared with the 0 μmol/L GCDC group, the apoptosis level, LDH release rate, and caspase-3 activity in the 50, 100, and 200 μmol/L GCDC groups gradually increased (all P<0.05) in a concentration-dependent manner. After RNAi-COX-2 lentivirus infection, COX-2 mRNA and protein expression levels in EBECs significantly decreased (all P<0.05). Compared with the GCDC group, the apoptosis rate of EBECs in the GCDC+ shCOX-2 group significantly decreased ( P<0.05). The GCDC+ shCOX-2 group showed increased cell proliferation activity, downregulated protein expressions of Bax, cleaved caspase-3, and p62, and upregulated protein expressions of Bcl-2, LC3 II/I, and Beclin-1 (all P<0.05). Conclusions:Inhibition of COX-2 improves GCDC-induced apoptosis of EBECs, and the mechanism may be related to the activation of the autophagy pathway.
