The Role Of The Pyroptosis In The Development Of Ulcerative Colitis

ObjectiveUlcerative colitis(UC)is a chronic non-specific intestinal inflammatory disease caused by various reasons.UC is characterized by repeated inflammation limited to the mucosal layer,usually starting from the rectum,which can extend continuously to the proximal colon,or throughout the colon over time.The main clinical manifestations include abdominal pain,diarrhea,tenesmus and mucous bloody stool,with or without iritis,anal fistula and other parenteral symptoms.The disease often presents the characteristics of recurrent attacks and remission,which is difficult to recover,so it not only affects the daily work and life of patients,but also easily induces UC-related colorectal cancer.With the rapid development of China’s economy and the changes of people’s lifestyle and diet habits,the number of UC patients is increasing.However,the etiology and pathogenesis of UC is currently unclear,and it has become the focus and difficulty of clinical UC research.As a newly discovered way of programmed cell death,Pyroptosis has been found to be widely involved in the occurrence and development of inflammatory diseases.At present,the exploration of the process of pyroptosis in inflammation and the application of pyroptosis in disease treatment have become research hotspots in molecular biology and cell biology.As the occurrence of pyroptosis and the release of inflammatory factors can directly affect the changes in physiology,morphology and pathology of cells and organisms.Therefore,an in-depth study of the relationship between UC and pyroptosis will is helpful for us to understand the role of pyroptosis in the occurrence and development of UC,and to provide new ideas and directions for clinical prevention of UC,screening of markers for UC patients,providing new targets for drug therapy and prognosis evaluation,which is of great significance in medical application.Method1.Construct UC model in vitro1.The CCK-8 experiment to detect the effect of different concentrations of DSS on human intestinal cancer cell line Caco-2.2.The CCK-8 experiment to detect the effect of different concentrations of DSS on human normal intestinal epithelial cell line NCM460.2.Cell morphology experiment1.Inverted microscope to observe the morphological changes of human intestinal cancer cell line Caco-2 in UC model.2.Inverted microscope to observe the morphological changes of of human normal intestinal epithelial cell line NCM460 in UC model.3.Flow Cytometry1.Annexin V-FITC/PI fluorescence staining to detect the apoptosis of human intestinal cancer cell line Caco-2 in UC model.2.Annexin V-FITC/PI fluorescence staining to detect the apoptosis of human normal intestinal epithelial cell line NCM460 in UC model.4.Western blot1.Western blot to detect the expression of pyroptosis-related proteins in human intestinal cancer cell line Caco-2 in UC model.2.Western blot to detect the expression of pyroptosis-related proteins in human normal intestinal epithelial cell line NCM460 in UC model.Result1.DSS has toxic effect on human intestinal cancer cell line Caco-2,and the toxic effect is concentration-dependent.2.DSS has a toxic effect on human normal intestinal epithelial cell line NCM460,and the toxic effect is concentration-dependent.3.human intestinal cancer cell line Caco-2 showes typical morphological changes of pyroptosis such as expansion,enlargement of volume,formation of large bubbles from plasma membrane and so on In UC model.4.human normal intestinal epithelial cell line NCM460 showes typical morphological changes of pyroptosis such as expansion,enlargement of volume,formation of large bubbles from plasma membrane and so on In UC model.5.The results of flow cytometry shows that there is membrane damage in human cancer cell line Caco-2 in UC model.6.The results of flow cytometry shows that there is membrane damage in human normal intestinal epithelial cell line NCM460 In UC model.7.The expression of pyroptosis-related proteins in human cancer cell line Caco-2 changes in UC model.8.The expression of pyroptosis-related proteins in human normal intestinal epithelial cell line NCM460 changes in UC model.Conclusion1.The cell model of ulcerative colitis in vitro was successfully constructed.2.Pyroptosis can occur in human intestinal epithelial cells in UC.3.Pyroptosis plays a positive role in the development of ulcerative colitis.