The Inhibitory Effects And Mechanisms Of Changtai In UC

Inflammatory bowel disease(IBD)is a non-specific chronic intestinal inflammatory diseases which comprises two primary forms of Crohn's disease(CD)and ulcerative colitis(UC).During it,ulcerative colitis is usually characterized clinically by bloody diarrhea,purulent mucus with stool and abdominal pain,etc.This kind of disease is typically recurrent and unknown etiology and pathogenesis till now,still,it is also accompanied with different complications.Therefore,ulcerative colitis is considered as one of refractory diseases by the World Health Organization(WHO).In our country,in recent decades the incidence of the disease has increased in recent decades and is prone to augment yearly.Traditional treatment methods cannot achieve a radical cure thoroughly,as a consequence,the patients who suffers this disease has a worse quality of survival.Due to ulcerative colitis is characterized mainly by inflammatory symptoms in colon tissue,recent researches suggest that strengthening the intestinal mucosal barrier function can help to reduce symptoms of the disease and relieve the pain.Therefore,inhibition of intestinal mucosal damage is becoming an effective treatment of recurrent of this disease.Compound Changtai is a total of 4 kind of compound preparation of traditional Chinese medicine,which is composed of cortex phellodendri,radix sanguisorbae,euphorbia humifusa and Polygonum flaccidum Meissn.The effective treatment of CD has been confirmed by a previous study which has proved that Changtai has anti-inflammatory and immune regulation mechanism effects.This study is committed to establish a proper animal model of UC to verify that the traditional Chinese medicine compound Changtai has equally effective regulating function on UC through the study of animal intestinal mucosal barrier function.Therefore,the main purpose of this study is designed to verify the compound Changtai that plays a significant a role in protecting the intestinal mucosal barrier function in ulcerative colitis,to explore the probably mechanisms through the results of proteomic analysis and to provide the drug treatment and theoretical foundation of clinical diseases such as UC.The methods: To design an experiment of ulcerative colitis by using dextran sodium sulfate(DSS).This experiment comprises six groups of animals,control group,model group,two different doses of Changtai treatment groups(4.3g/kg,17.1g/kg),Mesalazine group and Azathioprine group.Within the period of 7 days,mice should be administrated by gavage on time,and the body weight have to be monitored everyday.At the same time,the state of activity of each mice,loose stools and gross bleeding have to be recorded to calculate the disease activity index(DAI)of each animal;After mice sacrificed,the entire colon should be dissociated and observed,and the total length should be measured;observe inflammation of each group by hematoxylin and eosin staining;observe the ultrastructure of the colon by transmission electron microscopy;analyze the protein expression of tight junction proteins such as Occludin,Claudin-1 and ZO-1 by Western blot analysis;detect RNA expression of tight junction proteins such as Occludin,Claudin-1 and ZO-1 by Real-time PCR;assess mucosa cytokines levels of TNF-?,IL-6,IFN-? and IL-10 by ELISA kits;Non-standard quantitative proteomic LC-MS/MS method to detect the protein expression among control group,model group and high dose the protein expression of Chagntai group,in order to find differential proteins and proteomics analysis.The results:1?Compared to control group,the model group had significantly decreased in body weight,DAI,colon length after the DSS induced colitis(P < 0.05);on the contrary,high dose of Changtai had no significant difference with control group.Colon of model group was shortened obviously,congestion and edema,high dose of Changtai group showed mild inflammation such as oedema,a small amount of loose without congestion.2?HE staining results showed that inflammatory cell infiltrate of the model group was obviously observed.Meanwhile,high dose of Changtai group had no observed inflammation;3?TEM shows a damaged opening dot of the tight junction proteins in the colon of model group.Compared to the control group,the structure of the tight junction proteins of high dose of Changtai group is clear with no significant difference;4?Compared to high dose of Changtai group,the expression of protein of ZO-1,Occludin and Claudin-1 in model group has significantly decreased(P < 0.05).On the contrary,high dose of Changtai group shows no difference with the control group.5?Compared to high dose of Changtai group,the expression of RNA ofZO-1,Occludin and Claudin-1of model group has decreased,but did not show significant difference;6?Compared to high dose of Changtai group,the levels of TNF-?,IL-6,IFN-? and IL-10 of model group had decreased significantly(P < 0.05),On the contrary,high dose of Changtai group showed an opposite performance with the model group.7?proteomics analysis among differential proteins and pathways.This topic manifested that compound Changtai could effectively prevent the incidence of UC through the study of compound Changtai from protecting the intestinal mucosa barrier function in ulcerative colitis.The mechanism may be related to increasing the expression of tight junction proteins of intestinal epithelium and the regulation of intestinal immune function.The results had enlightened that we can further investigate the pathogenesis of UC from aspects of maintenance and strength of intestinal mucosal barrier.Understanding the effect of the physiopathology process of intestinal mucosa barrier in the pathogenesis of UC could also help to develop new treatments of UC,providing reference proof data for clinical treatments of UC.