Preventive And Therapeutic Effects Of Changqing Oral Liquid On Ulcerative Colitis And Its Mechanism

Intestinal mucous membrane plays a predominant role in hindering the invison of microorganism and toxin.During the ulcerative colitis (UC) period, damages at different levels, dysbacteriosis are showed in intestinal mucosa,which causes the invision of microorganism and toxin into intestinal tract,it would futher increase intestinal mucosa permeability and then induce or aggrevate intestinal inflammation.As a result of it, to amend the intestinal mucosa is conductive to inhibiting the inflammation of intestinal mucosa,which is a significant approach to improve UC. Bowel Clear Liquid,provided by South China Sea Institute of Oceanology Chinese Academy of Sciences.is a drug intended to application of patent.And there is no any related report on its efficacy in intestinal mucosa inflammation and influence in intestinal mucosa permeability.This study is to explore efficacy of Bowel clear liquid in♂Balb/c mice with TNBS-caused UC and its mechanism,which would provide a all-round pre-clinical basis for its clinical research.Objective:by applying the♂Balb/c mice with TNBS-caused UC,to study efficacy of Bowel Clear Liquid in TNBS-caused UC in♂Balb/c mice, and futher study its effect on intestinal mucosa of♂Balb/c mice and its mechanism.Methods:(1) The efficacy of Bowel Clear Liquid in TNBS-caused UC in♂Balb/c mice The mixture of5%trinitro-benzene-sulfonic acid and absolute ethyl alcohol(1:1) was injected with a dosage of150mg/kg into mice of all group except those of control group.All mice were randomly put into six groups,control group,model group, salazosulfapyridine group,and groups of low,middle and high dosage of Bowel Clear Liquid.The weight,faeces character and occult blood of each mouse were recorded each day, and on the basis of it,the disease activity index(DAI) of each mouse each day was got. Three days Iater,the mice were sacrificed,and the colon of each mouse was got, lesion level of colon was observed by naked eyes,and pathological grade was got.In conclusion,the DAI and UC of lesion level of colon in mice with TNBS-caused UC were observed to evaluate efficacy of Bowel Clear Liquid in treating UC. (2).Detection to the influence of Bowel Clear Liquid in intestinal mucosa permeability of mice with TNBS-caused UC The activity of diamine oxidase(DAO) of mice in all six groups was detected by Elisa to observe the influence of Bowel Clear Liquid in intestinal mucosa permeability.(3)Exploration on the mechanism of effect of Bowel Clear Liquid on intestinal mucosa permeability of mice with TNBS-caused UC. With approaches of RT-qPCR and Western Blot,the mRNA expression and protein expression of zo-l,occludin and claudin-1in mice of all six groups were explored,which is a mechanism study for the influence of Bowel Clear Liquid in intestinal mucosa permeability of mice with TNBS-caused UC.And with the Caco-2interfered with TNBS and then treated with Bowel Clear Liquid,the mRNA expression and protein expression of zo-1,occludin and claudin-1in Caco-2were detected too.(4) By the application of Caco-2to verify the mechanism for effect of xx on permeability of intestinal tractwith the establishment of Caco-2barrier model in vitro,to demonstrate the effect of Bowel Clear Liquid on permeability of Caco-2interfered by LPS.On the basis of chapter three,to further study the mechanism for effect of xx on permeability of intestinal tract through detecting the protein expression of claudin-1,zo-1and occludin in Caco-2.Results(1) The detection to efficay of Bowel Clear Liquid in treating♂Balb/c mice with TNBS-caused UC.Compared with model group, evident decrease in the DAI and lesion level of colon were observed in all three dosage groups of Bowel Clear Liquid,which else showed a dose-effect relationship.(2) Detection to the influence of Bowel Clear Liquid in plasm DAO activity of mice with TNBS-caused UC.Relative to model group,a signicant decline in plasm DAO activity in all other five groups was found.(3) As the result of RT-qPCR test,compared with model group a significantly raised mRNA expression of claudin-l,zo-1and occludin were showed in three dosage groups of Bowel Clear Liquid (P<0.05),and dose-effect relationship showed in this three groups was observed too.The result of approach of WB exhibited that relative to model group,a evidently raised protein expression of claudin-l,zo-1and occludin were detected (P<0.05)(4) with the establishment of barrier model in vitro using Caco-2,to demonstrate the michanism of the effect of Bowel Clear Liquid on permeability of intestinal tract.After the cultivation of Caco-2for20days, a arranement in alignment and tightness of Caco-2was observed,which was a model available for the study on permeability of intestinal tract in vitro.As the result of approach of western-blot showed, Bowel Clear Liquid could enhance the protein expression of claudin-l,zo-1and occludin in Caco-2(P<0.05)Conclusion:This research,with♂Balb/c mice with TNBS-caused UC,studied the efficacy of Bowel Clear Liquid in UC and its mechanism.A significant decline in DAI and relief in legion levels of mice were observed,which showed that Bowel Clear Liquid exhibited a significant efficacy in UC. Bowel Clear Liquid could obviously decrease DAO activity in plasm of mice with TNBS-caused UC,which indicated effect of Bowel Clear Liquid on UC was performed by influencing intestinal mucosa permeability. And Bowel Clear Liquid could enhance colon of mice with TNBS-caused UC and LPS-interfered Caco-2the mRNA and protein expression of zo-1,occludin and claudin-1,which indicated the influence of Bowel Clear Liquid in intestinal mucosa permeability might be through raising expression of zo-1,occludin and claudin-1.This study provides a pre-clinical basis for the clinical application.