Influence of dexmedetomidine on intestinal mucosal epithelial barrier function in spontaneous colitis mice
10.3760/cma.j.cn101480-20220111-00010
- VernacularTitle:右美托咪定对自发性结肠炎小鼠肠黏膜上皮屏障功能的影响
- Author:
Hailin XING
1
;
Haoran HU
;
Yong XIA
;
Honggang WANG
Author Information
1. 南京医科大学附属泰州人民医院（泰州市人民医院）麻醉科，泰州 225300
- Publication Type:Journal Article
- Keywords:
Colitis, spontaneous;
Dexmedetomidine;
Gene, interleukin-10;
Gene knockout;
Permeability;
Apoptosis;
Mice
- From:
Chinese Journal of Inflammatory Bowel Diseases
2022;06(3):240-247
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the influence of dexmedetomidine (DEX) on intestinal mucosal epithelial barrier function in spontaneous colitis mice.Methods:IL-10 gene knockout ( IL-10-/-) mouse was set as the model of spontaneous colitis. IL-10-/- mice were randomly divided into model group ( IL-10-/- group) and treatment group (DEX group) . The wild type mice (WT group) were set as control. The mice in DEX group was given intraperitoneal injection of 20 μg/kg DEX once a day for 14 days. The mice in IL-10-/- group and WT group were given the same volume of PBS intraperitoneal injection for 14 days. The disease activity index (DAI) , the weight change and colon histopathological score were caculated. The ratio of CD4 +CD45 + T cells in lamina propria of mucosa was detected by flow cytometry. The expression of inflammatory factors including interferon-γ (IFN-γ) and IL-17 was detected by enzyme linked immunosorbent assay (ELISA) . The mannitol permeability and transepithelial electrical resistance (TER) of colonic mucosa were detected by Ussing Chamber. The expression of tight junction proteins (Occludin and ZO-1) in intestinal epithelial cells was detected by confocal microscopy. The protein expression of Occludin, ZO-1, tumor necrosis factor-α (TNF-α) and tumor necrosis factor receptor 2 (TNFR2) was detected by Western blot. The colon cell apoptosis was detected by Tunel staining. The mRNA expression of TNF-α and TNFR2 was detected by fluorescence quantitative PCR. The differences of above indexes between the three groups were analyzed statistically. Results:(1) Inflammation: Compared with WT group, the DAI, histopathological score, ratio of CD4 +CD45 + T cells, expression of IFN-γ and IL-17 were higher in IL-10-/- group (all P<0.05) . Compared with IL-10-/- group, the DAI, histopathological score, ratio of CD4 +CD45 + T cells, expression of IFN-γ and IL-17 were lower in DEX group (all P<0.05) . (2) Permeability: Compared with WT group, the mannitol permeability was higher, TER was lower, the protein expression of Occludin and ZO-1 was lower in IL-10-/- group (all P<0.05) , and the location of proteins migrated. Compared with IL-10-/-group, the mannitol permeability was lower, TER was higher, the protein expression of Occludin and ZO-1 was higher in DEX group (all P<0.05) , and the location of proteins returned. (3) Apoptosis: Compared with WT group, the apoptosis cells were more, the protein expression of TNF-α and TNFR2 was higher, the mRNA expression of TNF-α and TNFR2 was higher in IL-10-/- group (all P<0.05) . Compared with IL-10-/- group, the apoptosis cells were less, the protein expression of TNF-α and TNFR2 was lower, the mRNA expression of TNF-α and TNFR2 was lower in DEX group (all P<0.05) . Conclusion:DEX is effective in ameliorating colitis of IL-10-/- mice, and the mechanism of improving intestinal mucosal barrier may be closely related to the reduction of intestinal permeability and the activity of TNF-α/TNFR2 apoptosis pathway.
