Effect And Mechanism Of Ento-A From Periplaneta Americana Extract On Chronic Recurrent Ulcerative Colitis

Objective:The therapeutic effect and mechanism of Ento-A extracted from Periplaneta americana on chronic recurrent UC were studied by flow cytometry,Western blot and HE methods.Methods:1.Establishment of DSS-induced chronic recurrent UC model in BALB/c mice,a certain number of mice were killed on 28th,42nd and 56th days respectively for CMDI and HS scores.The inflammatory degree of DSS model mice with different concentrations were evaluated,and the content of IL-1β in serum and IL-6 in colon tissue were determined by ELAS A method.2.Taking Olsalazine Sodium capsule as positive control,and different doses of Ento-A as tested drug,chronic recurrent UC mice were given corresponding drugs for 15 d by by means of intragastricadministration,normal control group and model control group were given NS solution respectively.The mice were killed the day after the last administration,the organ index was calculated,the colon length was measured,the CMDI and HS scores of mice colon were observed and recorded,the expression of pro-inflammatory cytokines in serum was determined by MSD method,and the expression of lymphocyte cytokines in spleen and mesenteric lymph nodes of mice in each group was detected by flow cytometry.The content of related proteins in colon mucosa was detected by WB method.Results:1.The DAI,CMDI and HS scores of chronic recurrent UC mice and the contents of IL-1β in serum and IL-6 in colon tissue were significantly increased(P<0.01).2.The experimental results of Ento-A in treating DSS-induced chronic recurrent UC mice showed that each dose group of Ento-A and Oshara Qin group can reduce DAI,CMDI,HS scores and colon index of mice,compared with the model control group.Ento-A can reduce the content of pro-inflammatory cytokines in serum,and there is no significant difference with Olsalazine Sodium capsule group.3.Flow cytometry results showed that Ento-A could effectively increase the content of IL-4 and Foxp3+in spleen lymphocytes and reduce the content of IFN-y and IL-17(P<0.05 or P<0.01).The contents of IL-4 and Foxp3+in mesenteric lymphnodes were decreased,but d IFN-γ and IL-17 were increased(P<0.05 or P<0.01).4.The results of Westen Blot showed that Ento-A could effectively reduce the expression of P-PI3K and P-AKT(P<0.05 or P<0.01)Conclusion1.Chronic recurrent UC animal model was induced by DSS,and the best modeling time was 49 days,and its concentration was 2.2%2.Ento-A from Periplaneta americana extract has a certain therapeutic effect on DSS-induced chronic recurrent UC in mice,which might be related to down-regulation of pro-inflammatory cytokines in serum of recurrent UC mice3.Ento-A could down-regulate the contents of IL-4 and Foxp3+ lymphocytes in mesenteric lymph nodes,up-regulate the expression of INF-γ and IL-17,and promote the balance of Th1/Th2 and Th17/Treg lymphocytes4.Ento-A from Periplaneta americana extract could treat DSS-induced chronic recurrent UC in mice by regulating PI3K/AKT pathway and inhibiting activation of P-PI3K,P-AKT and P-NF-κB.