Study On The Effects Of Total Flavones Of Ambrette On The Mice Model Of Crohn's Disease And Its Mechanisms Based On ERK1/2 And P38 MAPK

Objective:Crohn's disease(CD),one of the inflammatory bowel disease,is characterized by chronic non-specific intestinal inflammation,modern medical research has not yet confirmed its etiology.Patients with CD are usually difficult to get rid of drugs in their lives.Our preliminary study has confirmed that the Total Flavones of Ambrette(TFA)is effective against CD,but its specific mechanism is unclear.In this study,we established animal model of Crohn's disease with mice to explore the effcets of different dose of TFA on the model and further to investsgate the TFA's function upon the signal transduction of extracellular signal-regulated kinase 1/2(ERK1/2)and of p38 mitogen-activated protein kinase(p38MAPK).Methods:Female BALB/c mice were randomly divided into six groups,namely:normal control group,model group,SASP group,TFA low dose group,TFA middle dose group,TFA high dose group,model group,SASP group,TFA low dose group,TFA middle dose group,TFA high dose group establish model based on methods provided by the literature.Model establishment and administration at the same time.During the experiment,changes in disease activity index(DAI)and other indicators were recorded.At the end of the experiment,measure and weigh the colon tissue,observe colon gross morphological and pathological changes.Detect the expression of ERK1/2,phosphorylation-ERX1/2,p3 8MAPK,and phosphorylation-p3 8MAPK by using western blotting.Results:1.The difference of DAI between the groups was no statistically significant at the first week(P>0.05).The next week,the model group's DAI had a significant increase compared with the normal group(P<0.01);however,there were no significant differences in DAI between model group and treated groups(P>0.05).A week later,the DAI of SASP group and TFA middle dose group significantly decreased(P<0.05)and we couldn't see the differences between other treatment groups and model group.In the last week,the only group which showed a significant decrease compared to the model group was the TFA high dose group(P<0.01);there were no differences between other treatment groups and model group yet.2.The model group exhibited a significant increase in the weight length ratio of colon then normal group and treatment groups(P<0.05).Nonetheless,there was no differences among all the treated groups(P>0.05).3.Pathological scores of model group were significantly higher then control group(P<0.05),but there were no differences when compared to SASP group and TFA low dose group(P>0.05).However,pathological scores of TFA middle dose group and TFA high dose group showed a significant decrease when compared to model group(P<0.05).4.Protein levels of p38,P-p38,ERX1/2,P-ERX1/2 and tublin in the cytoplasm of colonic tissue of all groups were measured by using Western-blotting technique,contents of each proteins were presented by the gray value of corresponding stri.p at the same time.The p38MAPK and ERK1/2 protein were both expressed in the cytoplasm of normal rats.The model group showed a significant decrease in the protein level of ERK1/2(P<0.05)and a increase both in the level of P-EKR1/2.P-p38 and p38 levels compared to normal group(P<0.01,P<0.05).SASP group's protein level of ERK1/2 and P-ERK1/2 showed no significant difference compared to model group(P>0.05).At the same time,its expression of p38 and P-p38 showed a significant decrease when compared to model group(P<0.01).Protein levels of ERK1/2 and P-p38 in the TFA low dose group were lower than model group(P<0.01)while levels of P-ERK1/2 was higher than model group.Expression of TFA middle dose group's ERK1/2 and p38 had no significant difference with model group(P>0.05).However it's P-ERK1/2 and P-p38 decreased significantly compared with model group(P<0.05).TFA high dose group's protein level of ERK1/2 and P-ERK1/2 showed no significant difference compared to model group(P>0.05).However,its p38 and P-p38 levels were lower significantly than model group(P<0.01).Conclusions:TFA is effective in reducing disease activity index,pathological scores and weight length ratio,as well as controlling clinical symptoms.Decreasing P-ERK1/2 levels and down regulation of p38MAPK and P-p38MAPK in the colonic tissue may be possible mechanisms which are involved in the dose of TFA.