| Object In this study,D-IBS mouse model and chronic mild to moderate UC mouse model were established to observe the changes of TNF-α,IL-6 and IL-10 content in colonic mucosa of D-IBS model mice and UC model mice.2.The expression differences and histopathological changes of sodium-hydrogen transporter regulatory factor 4(NHERF4)and solute carrier 26A3(SLC26A3)in distal colonic mucosa of D-IBS model mice,UC model mice and healthy mice.By analyzing the difference of immuno-inflammatory factors and the expression changes of NHERF4 and SLC26A3 in intestinal mucosa of D-IBS and UC model mice,to explore whether mild to moderate UC and D-IBS have the same pathogenesis.Methods1.Construct the C57BL/6J mouse D-IBS model through the acute restraint experiment,observe the mouse’s body weight,intestinal bleeding,fecal water content,macroscopic tissue inflammation and damage of the colon,and histopathological findings under microscope,and obtain small samples at the same time.Mouse distal colon mucosal tissue.2.Construct a C57BL/6J mouse chronic mild-to-moderate colitis model with 1.5% oral DSS water solution,observe the mouse’s body weight,fecal water content,fecal occult blood,colonic inflammation damage and histopathological manifestations under microscope At the same time,the mouse distal colon mucosal tissue was obtained.According to Butzner JD standard for colon histology score,Sutherland LR standard for colitis disease activity index(DAI)score,and Gebose score for microscopic histological score.3.Enzyme-linked immunosorbent assay(ELISA)was used to measure the contents of TNF-α,IL-6 and IL-10 in the distal colon mucosa of D-IBS model mice,UC model mice and control mice.4.Observe the localization and expression of SLC26A3 and NHERF4 in intestinal mucosal epithelial cells by immunohistochemical methods.5.Western-blot method to observe the semi-quantitative expression of SLC26A3 and NHERF4 in the intestinal mucosa.Results1.In the C57BL/6J D-IBS mouse model test induced by the acute restraint experiment,the D-IBS model group mice have increased fecal water content,decreased body weight,and accelerated intestinal transit function similar to the clinical symptoms of human D-IBS.At the same time,its gross histology and histopathological changes are similar to those of human D-IBS.2.In the C57BL/6J mouse enteritis model test induced by a 1.5% concentration of DSS aqueous solution,the UC model group mice showed increased fecal water content and weight loss,which were similar to the clinical symptoms of human UC;and their DAI score,Gebose score,The gross histology and histopathological changes are similar to those of human UC.3.The content of TNF-α and IL-6 in the distal colon of the D-IBS model group and the UC model group are higher than the content of TNF-α and IL-6 in the distal colon of the normal control group.The content of IL-10 in the distal colon of the D-IBS model group and the UC model group was lower than that in the distal colon of the normal control group,and the difference was statistically significant.4.The expression of SLC26A3 protein in the intestinal mucosal tissue of mice in the D-IBS model group and the expression of SLC26A3 protein in the intestinal mucosal tissue of mice in the UC model group Were lower than the normal control group;the expression of NHERF4 protein in the intestinal mucosa of the D-IBS model group and the intestinal mucosa of the UC model group The expression of NHERF4 protein in tissues was higher than that of the normal control group.Conclusion1.The C57BL/6J D-IBS mouse model induced by acute restraint test and the C57BL/6J mouse enteritis model induced by low concentration of DSS are suitable to be used as animal models for the study of UC IBS and mild to moderate IBS.2.The activation of immune-inflammatory mechanism may be involved in the occurrence and development of D-IBS and UC.3.NHERF4/SLC26A3 may be involved in the occurrence and development of UC and D-IBS by changing the permeability of intestinal mucous layer. |
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