The Research On Observing The Effect Of Kuijieling Acting Mechanism In Ulcerative Colitis From NLPP3 Inflamasome Related Signal Pathway

Objectiveulcerative colitis(UC)represent major remitting and relapsing inflammatory disorders of the gastrointestinal tract and are characterized by chronic inflammation,abdominal pain,rectal bleeding,diarrhea and malnutritione.Recent animal studies suggested that the NLRP3 inflamasome is involved in the pathogensis of ulcerative colitis.The intracellular NOD-like receptor Nlrp3 is rapidly emerging as a crucial regulator of intestinal homeostasis.Although our previous studies showed the involvement of Th17 cytokines in the pathogenesis of UC.Contents1.The effect of Kuijieling on Weight of UC rats,the quality of colon length,colon,and pathological morphology?Inflammatory bowel disease,including ulcerative colitis and crohn's disease are two main types,it is because of the intestinal immune system disorders caused by a chronic,relapsing,persistent inflammatory diseases.Big rats UC model is set up,using and supervising and spirit treatment,observing rats colon length,quality,and pathological changes,in the form of evaluation and supervising and spirit for the treatment of UC rat induced by TNBS.Method60 SD rats were randomly divided into normal group,model group,Kuijieloing high dose group,Kuijieling medium dose group,Kuijieling low group and SASP group,10 in each group.In addition to the normal group,the rest of the group to trinitrobenzene sulfonic acid building 10 days,in the building after the observation of UC rats colon length,quality,and pathological changes of the form.ResultsIn plasma of rats,The length of the model group rats colon significantly shortened(P < 0.01);Collapse "spirit to provide dosage group and willow nitrogen cycling arsenic organism groups colon length increased significantly(P < 0.01).The quality of the colon model group was obviously increased(P < 0.01);Node spirit collapse spirit including dosage group and willow nitrogen cycling arsenic organism groups decrease of the quality of the colon(P < 0.05).Model group rats appear different degrees of weight loss,shorten the colon and colon quality increases,the pathological results show that the lamina propria disruption,bleeding,ulceration,blood capillary dilate,hyperemia,muscular layer and adventitia in a large number of acute or chronic inflammatory cell infiltration.Collapse and spirits high,medium and low dose group and willow nitrogen cycling arsenic organism group(0.5 g/kg)of the above model group significantly decrease relatively2.The Effect of Kuijieling on mRNA of NLRP3?Caspase-1?ASC in colonic mucosa of Ulcerative Colitis rats.The activation of NLRP3 regulation caspase 1 form called NLRP3 inflammatory signal complex body.It is composed of NLRP3,ASC and caspase 1 of three parts.By pathogens or dangerous mode makes the NLRP3 activation,makes the dependence on caspase 1 pro IL-1 beta,pro IL-18,pro IL-33 process occurs,which makes the form of the activity of these molecules to secrete and release.MethodMolding?grouping?dosing and drawing materials were the same as the first experiment.Colonic mucosa of rats,adequately homogenate,extracting total RNA.By fluorescence quantitative rt-pcr method in the detection of colonic mucosa of rats NLRP3,caspase 1,ASCmRNA content,and the results for statistical analysis.0 ul reaction system for reverse transcription.ResultsIn the model group rats colon mucosa NLRP3,caspase 1,ASC mRNA expression significantly higher(P < 0.01 or 0.05);Collapse and spirits high,medium and low dose group and willow nitrogen cycling arsenic organism group NLRP3,caspase 1,ASC mRNA expression were significantly reduced(P < 0.01 or 0.01).3.The effect of Kuijieling on the protein expression of NLRP3 ?Caspase-1?ASC in colonic mucosa of Ulcerative Colitis rats.To observe the effect of ulceration on the expression of NLRP3 in the colon mucosa of rats with ulcerative colitis.MethodAnimal models,groups and drugs were given to the same experiment.The total protein of colonic mucosa was extracted and the protein content was determined by BCA method.Western blot was used to detect the relative expression of NLRP3,caspase-1 and ASC protein in the colon mucosa of rats,and the results were statistically analyzed.ResultsModel group rats NLRP3?Caspase-1and ASC protein expression is higher than normal group(P < 0.05),and collapse spirit high,medium and low dose group of NLRP3?Caspase-1? ASC and SASP group expression is lower than the model group(P < 0.05 or P < 0.01).4.The effect of Kuijieling on the IL-1? ?IL-18?IL-33 in colonic mucosa of Ulcerative Colitis rats.NLRP3 activated,its oligomerization makes its PYD domain structure combined with ASC PYD structure domain,and through the CARD area raise activation makes Caspase procaspase-1,the activation of Caspase-1 to pro IL-1 beta? pro-IL-18 ?pro-IL-33 to shear,make its have the biological activity of mature processing form before cytokines to participate in the intestinal immune response and inflammatory process,they are in ulcerative colitis plays a key role in the occurrence and development process of NLRP3 activation,the oligomerization makes the PYD domain structure combined with ASC PYD structure domain,and through the CARD area raise activation makes Caspase procaspase-1,the activation of Caspase-1 to pro-IL-1 beta,pro-IL-18 ?pro-IL-33 to shear,make its have the biological activity of mature processing form before cytokines to participate in the intestinal immune response and inflammatory process,they are in the process of the occurrence and development of the ulcerative colitis plays a key role.MethodAnimal models,groups and drugs were given to the same experiment.The contents of IL-1,IL-18 and IL-33 in the colon mucosa of rats were detected by enzyme-linked immunosorbent assay.ResultsIn the model group rats colon mucosa IL-1 beta,IL-18,the content of IL-33 was obviously higher than that of normal group(P < 0.01),and collapse spirit high,medium and low dose group and SASP group in colonic mucosa IL-1 beta,IL-18,IL-33 content is significantly lower than the model group(P <0.05 or P < 0.01);ConclutionThe result of the study demonstrate that,the activation of NLRP3 inflammasome as well as the related inflamatory factors may be relevant to the intestinal inflammasome of rats.Kuijieling is able to alliviate the chronic ulcerrative colitis by inhabiting the NLRP3 inflammasome.