A Vitro Study On Biology Behavior Of Colon Dendritic Cell Of Rats With Irritable Bowel Syndrome Model

Objective: We are for the purpose of studying the correlation among the Runx3, TGF-β1 and dendritic cells in the paroxysm of IBS and it provides clues to the pathogenesis of IBS.Methods:We set up the model of IBS through the method of intracolonic instillation of acetic acid and restraint stress. The morphology of dendritic cells in a piece of colon obtained from each rats were observed through transmission electron microscope(TEM). Afterwards, the dendritic cell of the rat colon mucosa lamina propria isolated by the means of density gradient centrifugation was cultivated in vitro for 12 days and chosen by MACS Separators. Then RUNX3 short hairpin RNAs(sh RNA-RUNX3) were transfected by si RNA technology and the dendritic cells were isolated. Finally, after the cells were cultivated for 7 days, the DC phenotype detection was performed by the flow cytometry and the expression of TGF-β1 and Runx3 protein were tested by western blotting.Results: 1. Under different expansion capacity, the AWR score and the electroactivity of abdominal muscle were compared between the model group and normal group. And the model group has remarkably higher AWR score. It demonstrates that the sensibility of viscera has enhanced. 2.The expression rate of OX62 on the surface of peyer’s pathes in the IBS model group is obviously higher than that of normal group. 3. The dendritic cell observed by inverted fluorescence mirror is accord with the structural characteristics of DC.4. The OX62, CD80, CD86, MHC-II on the surface of DC of normal group have lower expression rate than that of IBS group. However, there is no difference between transfection group and non-transfection group.5. The results of western blotting demonstrate that the expression amount of Runx3 protean in transfection group has remarkably reduced than that of non-transfection group. And there is no obvious difference between the TGF-β1 expression rate of transfection group and non-transfection group.Conclusions: 1. Rats model of IBS by intracolonic instillation of acid and restraint stress and single restraint stress methods are accord with visceral hypersensitivity of IBS, this is practicable for experimental study.2. The increase of abnormal dendritic cells might influence the occurrence and development of irritable bowel syndrome patients.3. Runx3 and TGF- β1 needs to be further studies in irritable bowel syndrome patients.4. TGF-β1 signal pathway might not be involved in Runx3-regulated maturation of dendritic cells in irritable bowel syndrome patients.