The Effect Of Kuejieling On Caspase-1 Mediated Pyroptosis In Ulcerative Colitis Rats

Objective:Ulcerative colitis(UC)is a kind of chronic non-specific inflammatory disease involving the colorectal,and it is a part of the inflammatory bowel disease(IBD).Pyroptosis is a newly discovered programmed cell death related to inflammation,which is closely related to infectious diseases and autoimmune diseases.The pore-forming protein gasdermin D(GSDMD)plays an important role in the process of cell pyroptosis.After being cutted by caspase-1,GSDMD-N terminal domain forms small pores on the cell membrane,and then promotes the release of inflammatory cytokines,and eventually leads to pyroptosis.Previous experimental studies have explored that the effect of kuijieling on the NF ?B pathway and NLRP3 inflammasome.Therefore,the aim of this study is to explore the regulation effect of caspase-1 mediated pyroptosis,and deeply reveal the mechanism of action and and targets in the treatment of UC by kuijieling.Methods:Male SPF SD rats were randomly divided into normal group,model group,Kui jieling high-dose group,kui jieling medium-dose group,kui jieling low-dose group and SASP group.Except the normal group,the other five groups were induced into UC model by 2,4,6-trinitrobenzenesulfonic acid(TNBS).After 3 days,drug intervention was given to each group for 10 days except the normal group and model group.During the experiments,the general condition of the rats and the change of body weight were recorded.After the animal experiment,we measured the length of the colon and the weight of the colon,and collected the colonic mucosa of the rats.The mRNA expression of IL-1? and IL-18 in the colonic mucosa were detected by real-time fluorescent quantitative PCR.The protein expression of GSDMD,Caspase-1,IL-1? and IL-18 in were detected by Western Blot.Results:1?Compared with the normal group,model group rats presented weight lost(P=0.001),the length of colon shortened(P=0.000)and the weight of colon increased(P=0.002).Compared with the model group,the weight of rats in kui jieling high-dose group and SASP group were increased significantly(P=0.004,0.003);in kuijieling high-dose group,kuijieling medium-dose group and SASP group,the length of colon was increased significantly(P=0.030,0.024,0.023);In kuijieling high-dose group,kuijieling medium-dose group,kuijieling low-dose group and SASP group,the weight of colon was lighter than the model group(P=0.007,0.012,0.01,0.017).2?Compared with the normal group,the mRNA expression of IL-1? in the model group was significantly increased(P=0.000).Compared with the model group,the mRNA expression of IL-1? in kuijieling high-dose group and SASP group were decreased significantly(P=0.005,0.010).Compared with the normal group,the mRNA expression of IL-18 in the model group was significantly increased(P=0.016).Compared with the model group,the mRNA expression of IL-18 in kuijieling high-dose group and SASP group were decreased significantly(P=0.039,0.025).3?Compared with the normal group,the protein expression of GSDMD in the model group was significantly increased(P=0.004).Compared with the model group,the protein expression of GSDMD in kuijieling high-dose group and SASP group were decreased significantly(P=0.016,0.025).Compared with the normal group,the protein expression of caspase-1 in the model group was significantly increased(P=0.004).Compared with the model group,the protein expression of caspase-1 in kuijieling high-dose group and SASP group were decreased significantly P=0.016,0.037).Compared with the normal group,the protein expression of IL-1? in the model group was significantly increased(P=0.001).Compared with the model group,the protein expression of IL-1? in kuijieling high-dose group,kuijieling medium-dose group and SASP group were decreased significantly(P=0.008,0.034,0.025).Compared with the normal group,the protein expression of IL-18 in the model group was significantly increased(P=0.000).Compared with the model group,the protein expression of IL-18 in kui jieling high-dose group and SASP group were decreased significantly(P=0.007,0.012).Conclusion:According to the comprehensive experimental results,there may beaclose relationship between the inflammation in UC rats and pyroptosis.We can make a conclusion that kuijieling has a therapeutic on UC rat models,and the mechanism may be related to its inhibition of caspase-1 activation,further inhibition of GSDMD activation,reduction of cell pyrosis,and reduction of il-1 and 1l-18 production and release.