Experimental Study On Establishment Of Ibd Animal Models And Tetramethylpyrazine Intervention

Purpose: IBD contains CD and UC,and it is a kind of chronic and non-spesific intestinal inflammatory disease which is widely spread in China and its cause is still unknown.We are purposed to establish animal models of CD and UC respectively,to find out the differences between the two animal models and then treat these animals with tetramethylpyrazine(TMA)to study whether this medicine would be a cure of IBD and explore the probable molecular mechanism.Methods: to establish TNBS induced CD rat model and oxazolone induced UC mouse model respectively and which were appraised by the damage grade of the intestines and the rate of Th1 cells and Th2 cells through FCM.The two animal models were verified by DNA array and Western Blot.SMC,PBMC and LPMC from the animals of CD model and UC model were sorted and cultured with gradient concentration of TMP,the rate of Th1 cells and Th2 cells were detected through FCM,the apoptosis rate of these cells were detected by Annexin V kit,the expression of IL-4,IL-5,IL-10,IL-13,IL-33,VCAM-1,IL-2,IL-12,IFN-?,TNF-?,ICAM-1,i NOS,NF-?B,NF-AT,AP-1 and P-selectin were detected by RT-PCR.Results: CD rat model and UC mouse model were successfully established,and the protocol of CD rat model was updated.The UC mouse model was a model of Th2 cells dominated disorder of immunity which is close to IBD of human being,while the CD rat model was more like a model of Th1 cells activated inflammation.Compared to normal rats,there were up-regulated expression of TSC-22,STAT3,IFIM1,PAI-2 and down-regulated expression of Galectin-9 and MIP-1a in CD rat model.Compared to normal mice,there were up-regulated expression of Ig HG3,PAI-2,NOS2 A and down-regulated expression of MDR-1,HGF and PXR in UC mouse model.2mg/ml TMA could significantly suppress the expression of P-selectin and promote the apoptosis of LPMC as well.TMA could also inhibit the apoptosis of histocyte of intestine and down-regulate the expression of IL-4,IL-5,IL-10,IL-13,IL-33,VCAM-1,IL-2,IL-12,IFN-?,TNF-?,ICAM-1and iNOS.Conclusion: TMA could induce the apoptosis of histocyte of intestine and promote the apoptosis of LPMC,TMA also down-regulate the expression of inflammatory cytokines such as IL-4,IL-5,IL-10,IL-13,IL-33,VCAM-1,IL-2,IL-12,IFN-?,TNF-?,ICAM-1,i NOS and P-selectin.TMA may play an important role in the treatment of IBD in the future.