Effects of gene silence on immune function of colonic dendritic cells in rats with irritable bowel syndrome visceral hypersensitivity
10.3760/cma.j.issn.0254-1432.2016.02.005
- VernacularTitle:基因静默对肠易激综合征内脏高敏感大鼠肠道树突状细胞免疫功能的影响
- Author:
Zhaomeng ZHUANG
;
Lu ZHANG
;
Liyuan TAO
;
Meng LI
;
Bin LYU
- Publication Type:Journal Article
- Keywords:
Protein disulfide isomrase A3;
Dendritic cells;
Interleukin-4;
Interleukin-9;
CD4-positive T-lymphocytes;
CD8-positive T-lymphocytes;
Visceral hypersensitivity
- From:
Chinese Journal of Digestion
2016;36(2):96-100
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the role and mechanism of protein disulfide isomerase A3 (PDIA3) in abnormal colonic mucosa immune response of irritable bowel syndrome (IBS) rats with visceral hypersensitivity.Methods A total of 48 SD rats were divided into blank control group (n=12),empty virus group (IBS-1) (n=12),PDIA3 silencing group (IBS-2) (n=12) and model control group(IBS-3) (n=12).The expression of CD103 in ileocecus colonic tissues was detected by immunohistochemistry.Dendritic cells (D C) of mesenteric lymph node were isolated by flow cytometry.CD4+/CD8+ T cells of spleen were separated by immune-magnetic beads sorting technique.DC and CD4+/CD8+ T cells were co-cultured.The proliferation ability of lymphocytes promoted by DC was measured by methyl thiazolyl tetrazolium (MTT).The secretion levels of interlcukin (IL) 4 and IL-9 of CD4+/CD8+ T cells stimulated by DC were determined by enzyme linked immunosorbent assay (ELISA) method.Independent sample t-test was performed for statistical analysis.Results The cell number of CD103 positive DC of rats colon of blank control group,IBS-3 group was 6.25±1.14 and 10.83± 1.03(t=10.07,P＜0.05);that of IBS-2 group was 7.42 ± 0.90,and compared with that of IBS-3 group,the difference was statistically significant (t=-9.25,P ＜ 0.05).The MTT value of CD4+ T cells proliferation stimulated by DC of blank control group and IBm3 group was 0.54±0.01 and 0.60±0.01 (t=3.373,P＜0.05);that of IBS-2 group was 0.53±0.01,and compared with that of IBS-3 group,the difference was statistically significant (t =-3.139,P ＜ 0.05).The MTT value of CD8+ T cells proliferation stimulated by DC was 0.52±0.01 and 0.59±0.00(t=3.539,P＜0.01);that of IBS-2 group was 0.54±0.01,and compared with that of IBS-3 group,the difference was statistically significant (t=-3.183,P＜0.05).The level of IL-4 secreted by CD4+T cells promoted by DC of blank control group and IBS-3 group was 10.24±0.09 and 16.61±1.00 (t=3.222,P＜0.05);that of IBS-2 group was 11.75±0.54,and compared with that of IBS-3 group,the difference was statistically significant (t=-3.539,P＜0.01).The level of IL-9 secreted by CD4+T cells stimulated by DC of blank control group and IBS-3 group was 15.86±10.19 and 43.51±11.32 (t=4.529,P＜0.05);that of IBS-2 group was 29.05±2.09,and compared with that of IBS-3 group,the difference was statistically significant (t=-6.841,P＜0.01).The level of IL-4 secreted by CD8+T cells promoted by DC of blank control group and IBS-3 group was 7.35±0.12 and 13.91±0.57 (t=19.557,P＜0.01);that of IBS-2 group was 8.63± 0.24,and compared with that of IBS-3 group,the difference was statistically significant (t =-14.782,P＜0.01).The level of IL-9 secreted by CD8+T cells stimulated by DC of blank control group and IBS-3 group was 29.12±5.14 and 60.70±11.02 (t=4.122,P＜0.05);that of IBS-2 group was 37.17±2.65,and compared with that of IBS-3 group,the difference was statistically significant (t=-3.255,P＜ 0.05).Conclusions Protein disulfide isomerase A3 may mediate the process of DC activating T lymphocyte levels of related cytokines,cause abnormal immune response of colonic mucosa and promote the genesis of IBS visceral hypersensitivity.
