Effect of Hypoxia-induced Factor 1α and 2α in Ethanol-induced Intestinal Epithelium Cellular Barrier Dysfunction
10.11969/j.issn.1673-548X.2024.10.023
- VernacularTitle:HIF对乙醇诱导肠上皮细胞屏障功能损伤的影响
- Author:
Yibing HU
1
;
Yafang LI
;
Chong LU
Author Information
1. 321000 浙江大学医学院附属金华医院消化内科
- Keywords:
Hypoxia-induced factor;
Ethanol;
Intestinal epithelium cellular barrier
- From:
Journal of Medical Research
2024;53(10):129-134
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of hypoxia-induced factor 1α and 2α(HIF-1α and HIF-2α)on ethanol-in-duced intestinal epithelium cellular barrier dysfunction of colorectal adenocarcinoma cell(Caco-2)monolayers and its mechanism.Methods Intestinal epithelium monolayer cellular barrier model was obtained by Caco-2 in vitro and induced by ethanol with different concentrations.The cell viability was measured by MTT method.Caco-2 cells were treated with or without ethanol according to the opti-mum concentration and time.The secretions of interleukin 1 β(IL-1 β)and interleukin 6(IL-6)were detected by ELISA.The expres-sions of HIF-lα,HIF-2α,and Neu3 were analyzed by Western blot and RT-PCR.The permeability of Caco-2 cell monolayers was evaluated by transepithelial electrical resistance(TEER).Next,Caco-2 cells were transfected with small interfering RNA(siRNA)to knock down the expression of HIF-1α and HIF-2α.The cell viability,the levels of IL-1 β and IL-6,and the expression of Neu3 and TEER in each group were detected,respectively.Then,the expression of Neu3 was inhibited with siRNA to assess the permeability of Ca-co-2 cell monolayers by TEER.Results The inhibitory effect of ethanol on Caco-2 cells was dose-dependent when the concentration of ethanol was higher than 5％.When Caco-2 cells were treated with ethanol with a concentration of 5％for one hour,the levels of proinflammatory cytokines IL-1 β and IL-6 were promoted.Compared to the control group,ethanol also induced the expression of HIF-1α,HIF-2α,Neu3 and reduced TEER values(P＜0.05).After HIF-1α or HIF-2α siRNA interference in ethanol-induced Caco-2 cells,the cell viability was inhibited,the secretions of IL-1 β and IL-6 were significantly elevated and the values of TEER were decreased(P＜0.05).Interestingly,the expression of Neu3 was inhibited when HIF-2α was deleted but not HIF-1α(P＜0.05).Furthermore,TEER analysis showed an increased cellular permeability when the expression of Neu3 was inhibited(P＜0.05).Conclusion The HIF-1α or HIF-2α signal pathway may be involved regulation process of ethanol-induced increased permeability of the intestinal epithelium cellular barrier.In this process,Neu3 may be mediated by HIF-2α but not HIF-1α,contributing to intestinal epithelium cellular barrier dysfunction in ethanol-induced Caco-2.
