To explore the effect of A2AR on endotoxin-induced inflammatory injury of Caco-2 intestinal epithelial cells from the perspective of autophagy
10.19405/j.cnki.issn1000-1492.2024.09.019
- VernacularTitle:基于自噬角度探究A2AR对内毒素诱导的Caco-2肠上皮细胞炎性损伤的影响
- Author:
Yu CHEN
1
;
Xiaoji WANG
;
He LI
Author Information
1. 海南医学院第一附属医院普通外科,海口 570000
- Keywords:
adenosine A2A receptor;
Caco-2 intestinal epithelial cells;
inflammatory bowel disease;
lipopolysac-charide;
cell damage;
autophagy
- From:
Acta Universitatis Medicinalis Anhui
2024;59(9):1636-1642
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect and mechanism of adenosine A2A receptor(A2AR)on lipopolysac-charide(LPS)-induced inflammatory injury of Caco-2 intestinal epithelial cells.Methods Caco-2 cells were di-vided into control group,LPS group(treated with 10 μg/ml LPS for 12 h),A2AR agonist(CGS21680)group(pretreated with 50 nmol/L CGS21680 for 10 min),CGS21680+LPS group(pretreated with 50 nmol/L CGS21680 for 10 min,treated with 10 μg/ml LPS for 12 h),cell viability was determined using CCK-8 assay,the secretion levels of tumor necrosis factor-α(TNF-α),interleukin-1 β(IL-1 β)and interleukin-6(IL-6)in cell su-pernatant of each group were determined using ELISA.mRNA expression levels of TNF-α,IL-1 β and IL-6 in cells of each group were detected by real-time fluorescence quantitative PCR,the protein expression levels of microtubule associated light chain protein 3(LC3)-Ⅱ/LC3-Ⅰ and autophagy associated protein(Beclin1)in cells of each group were analyzed using Western blot analysis.Caco-2 cells were then divided into control group,LPS group(pretreated with 50 nmol/L CGS21680 for 10 min),CGS21680+LPS group(pretreated with 50 nmol/L CGS21680 for 10 min,treated with 10 μg/ml LPS for 12 h),CGS21680+LPS+Rapa group(pretreated with 50 nmol/L CGS21680 for 10 min,treated with 10 μg/ml LPS and 5 μmol/L Rapa for 12 h),cell viability was deter-mined using CCK-8 assay,the secretion levels of TNF-α,IL-1 β and IL-6 in cell supernatant of each group were determined using ELISA.Results Compared with the control group,the viability of Caco-2 cells in LPS group sig-nificantly decreased(P＜0.05),the levels of TNF-α,IL-1 β and IL-6 in supernatant significantly increased(P＜0.05),the mRNA relative expressions of TNF-α,IL-1β,IL-6 in cells significantly increased(P＜0.05),the LC3-Ⅱ/LC3-Ⅰ ratio and the relative expression of Beclin1 protein were significantly up-regulated(P＜0.05).Compared with LPS group,the viability of Caco-2 cells in CGS21680+LPS group significantly increased(P＜0.05),the levels of TNF-α,IL-1 β and IL-6 in supernatant significantly decreased(P＜0.05),the mRNA rela-tive expression levels of TNF-α,IL-1β,IL-6 in cells were significantly down-regulated(P＜0.05),and the ratio of LC3-Ⅱ/LC3-Ⅰ and the relative expression of Beclin1 protein were significantly down-regulated(P＜0.05).In addition,compared with CGS21680+LPS group,the viability of Caco-2 cells in CGS21680+LPS+Rapa group significantly decreased(P＜0.05),and the levels of TNF-α,IL-1 β and IL-6 in the supernatant also significantly increased(P＜0.05).Conclusion A2 AR agonist can reduce the inflammatory injury of Caco-2 intestinal epithe-lial cells induced by LPS and improve cell viability,which may be related to its inhibition of autophagy.
- Full text:2024122421193652956基于自噬角度探究A2AR对...2肠上皮细胞炎性损伤的影响_陈禺.pdf
