| Objective To construct a mouse model of resveratrol in the repair of inflammatory bowel disease,and to detect the expression of SUMO1 and its related signaling pathways in colorectal mucosa of mice in negative control group,IBD model group and IBD group,and to explore the role and mechanism of resveratrol in the regulation of SUMO1 in the repair of inflammatory bowel disease.Methods 1.Establishment of IBD model in mice repaired with resveratrol Male BALB/c mice aged 6 weeks(about 20g)were selected to construct the IBD model group(model group),resveratrol treatment group(experimental group)and negative control group induced by dextran sulfate sodium(DSS).Mice in the model group continued to drink double-steamed water containing 3% DSS,while mice in the experimental group continued to drink double-steamed water containing 3% DSS and were given resveratrol(50mg/kg)daily,while mice in the negative control group drank double-steamed water.The weight of mice was weighed at the prescribed time point every day and the fecal characteristics of mice were observed.When the positive mice were found to lose weight and fecal bleeding(stool bleeding)and other colorectal inflammation symptoms,it indicated that the model of inflammatory bowel disease was successfully constructed.Three groups of mice were killed by inhalation of excessive carbon dioxide.Colonic and rectal mucosal tissues and spleen tissues of three groups of mice were taken for further detection.2.Hematoxylin-eosin staining(HE)and immunohistochemistry(IHC)HE and IHC were used to observe the structure of colorectal mucosa and spleen tissues of three groups of mice and detect the expression level of proliferating cell nuclear antigen(PCNA)in colorectal mucosa of three groups of mice.3.Detection the expression of SUMO1,Wnt5 a,?-catenin and related inflammatory factors in colorectal mucosa and spleen in tissuesReal-time quantitative polymerase chain reaction(PCR),Western-blot and IHC were used to detect the expression of SUMO1,Wnt5 a,beta-catenin and related inflammatory factors in colorectal mucosa and spleen tissues of three groups of mice.4.To detect the expression of SUMO1 and beta-catenin in intestinal tissues of patients with inflammatory bowel disease and colorectal cancer.The expressions of SUMO1 and beta-catenin in colorectal tissues of patients with inflammatory bowel disease and colorectal cancer and normal persons were detected by immunohistochemistry.Results 1.Establishment of IBD model in mice repaired with resveratrol The results showed that from the 7th day of modeling,the weight of model group mice continued to decline,while that of negative control group mice increased steadily,while that of experimental group mice remained stable at a certain level and increased slightly.When observing the obvious hematochezia in the model group,the hematochezia in the experimental group was lighter and the hematochezia time was later than that in the model group,which indicated that resveratrol was successfully used to repair the inflammatory bowel disease in mice.The disease activity index(DAI)of mice in the model group increased significantly from the 7th day.The DAI of mice in the experimental group increased slightly from the 7th day and remained unchanged at a certain level.The DAI of mice in the negative control group continued to be 0.Three groups of mice were killed by carbon dioxide excessive inhalation.Colon and rectum mucosa and spleen tissues were taken from each group,and the length of colon and rectum in the experimental group was longer than that in the model group.HE staining showed that the structure of colon and rectum mucosa and spleen in the experimental group was more orderly than that in the model group,and the infiltrating inflammatory cells were significantly reduced.The results showed that the expression level of PCNA in colorectal mucosa of experimental mice was significantly higher than that of model mice.2.To detect the expression of SUMO1,Wnt5 a,beta-catenin and related inflammatory factors in colorectal mucosa and spleen tissues.The expression of SUMO1,Wnt5 a,beta-catenin and related inflammatory factors in colorectal mucosa and spleen of mice in negative control group,model group and experimental group were detected by real-time quantitative PCR,Western-blot and immunohistochemistry.Real-time quantitative PCR results showed that the expression levels of SUMO1,IL-6,IL-8,TNF-a and IL-1beta in colorectal mucosa and spleen tissue of mice in experimental group were significantly lower than those in model group,and the expression levels of IL-10 and SENP1 in colorectal mucosa of mice in experimental group were significantly higher than those in model group;the expression levels of IL-10 and SENP1 in spleen tissue of mice in experimental group were significantly higher than those in model group.Group.Western-blot results showed that the expression levels of SUMO1,Wnt5 a,beta-catenin and NF-kappa B in colorectal mucosa of mice in the experimental group were significantly lower than those in the model group.3.To detect the expression of SUMO1 and beta-catenin in colorectal inflammatory tissues of patients with enteritis and colon cancer.The expression levels of SUMO1 and beta-catenin in colorectal mucosa of patients with inflammatory bowel disease and colorectal cancer and healthy control group were detected by IHC.The results showed that the expression levels of SUMO1 and beta-catenin in colorectal mucosa of patients with inflammatory bowel disease and colorectal cancer were significantly higher than those of normal control group.Conclusions In this study,we successfully established a model of resveratrol-induced inflammatory bowel disease in mice.The expression level of SUMO1 in the colorectal mucosa and spleen of the experimental group was significantly lower than that of the model group,accompanied by decreased expression of inflammatory factors and decreased activation of Wnt/beta-catenin and NF-kappa B signaling pathways.The expression of SUMO1 in intestinal lesions of patients with inflammatory bowel disease and colorectal cancer was significantly higher than that of normal control group,suggesting that SUMO1 can promote inflammatory bowel disease and colorectal cancer.These results suggest that SUMO1 plays an important role in the repair of inflammatory bowel disease in mice by resveratrol,which suggests that resveratrol may repair inflammatory bowel disease by down-regulating the expression of SUMO1.This study provides theoretical and experimental basis for clinical application of resveratrol in the treatment of inflammatory bowel disease. |
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