Effect Of The Enema Treatment Of Yu’s Clearing Heat And Activating Blood On β-arrestin 2 Signaling Pathway In Ulcerative Colitis

Objective To establish a model of ulcerative colitis in Spraque-Dawley(SD)rats by preserving clystering trinitrobenzene sulfonic acid(TNBS)/ethanol.Observing the effect of the treatment of clearing heat and activating blood on healing the rats with ulcerative colitis and the influence on β-arrestin 2 signaling pathway.exploring possible mechanism of the prevention and treatment of ulcerative colitis.Methods 60 healthy male SD rats were randomly divided into normal group,model group,mesalamine group,the Chinese medicine low,medium and high dose groups.Normal group were clystered 0.85 ml of distilled water,the rest were clystered 0.85 ml trinitrobenzene sulfonic acid/ethanol mixed solution.Then we start drug intervention,clyster the low,medium and high dose group with the 2ml traditional Chinese medicine whose concentration is2.1g/ml,4.2g/ml,8.4g/mldifferently.Clyster the mesalamine group with 2ml of 0.03g/ml mesalamine suspension fluid,the normal group and model group were clystered with distilled water every day.10 days after the treatment,narcotize the rats,intercept their colon tissue after death,observe the condition of inflammation and ulcers with a magnifying glass,and take a small piece of colon tissue,fixed in 10% formalin solution,paraffin embedding,sectioning.Used for hematoxylin-eosin dye(HE)staining,and immunohistochemical and histological observation and diagnosis of β-arrestin2,NF-κB protein.Results(1)General condition of rats: normal group rats’ general condition was well,the nature of the stool and the number of times have not seen obvious change;Model group rats’ general condition was bad,emaciation,diarrhea,mucous purulent blood stool,slow action;The drug intervention groups were better than the model group.(2)Colon tissue morphological grading: Compared with the normal group,the rest five groups’ scores were increased significantly(P<0.01);Compared with model group,the drug intervention groups’ scores were lower(P<0.05),except Chinese medicine low dose group decreased significantly(P<0.01).The mesalamine group and the Chinese medicine groups have no differences.(3)HE dyeing observation and pathological damage rating: the basic structure of the colon tissue in rat normal group was normal.The model group’s colon tissue was damaged severly,drug intervention groups’ colon’s injury was significantly reduced than the model group.Compared with the normal group,the rest five groups’ scores were increased significantly(P<0.01);Compared with model group,the drug intervention groups’ scores decreased significantly(P<0.01),except the mesalamine group(P<0.05).The mesalamine group and the Chinese medicine groups have no differences.(4)β-arrestin2,NF-κB protein immunohistochemical expression: Compared with the normal group,the rest groups’ expression of β-arrestin2 protein was decreased,the mesalamine group decreased with statistically significant(P<0.05)and the model group decreased significantly(P<0.01).Compared with model group,the mesalamine group and the Chinese medicine low dose group increased with statistically signigicant(P<0.05),.The other two groups of Chinese medicine show significant difference in increase ofβ-arrestin2(P<0.01).The mesalamine group and the Chinese medicine groups have no differences.Compared with the normal group,the model groups’ expression of NF-κB protein increased(P<0.01),the Chinese medicine middle dose group decreased with statistically signigicant(P<0.05),the others showed no differences.Compared with model group,the mesalamine group and the Chinese medicine groups show significant difference in decrease of NF-κB(P<0.01).The mesalamine group and the Chinese medicine groups have no differences.Conclusion(1)By clystering rats with trinitrobenzene sulfonic acid/ethanol can successfully induced ulcerative colitis model rats,which conforms to the human’s pathogenesis of ulcerative colitis.(2)The treatment of Yu’s clearing heat and activating blood can improve the general situation of rat with ulcerative colitis,and reduce colon injury as well as the pathological tissue damage,can relieve the damage from inflammation.(3)The treatment of clearing heat and activating blood can suppress the fuction of NF-κB by upregulating the expression of β-arrestin2 to relieve the inflammation.