Objective:This study used Huaihua San in the treatment of ulcerative colitis patients who were diagnosis by large intestinal damp-heat in TCM,to observe and compare the changes before and after treatment in patients with symptoms,signs,comprehensive changes in symptoms,blood related index,improved the Mayo total score,overall syndrome effect,clinical comprehensive efficacy,to evaluate the clinical effect of Huaihua San in the treatment of UC large intestinal damp-heat syndrome patients.Through the replication of UC model in rats,the effects of Huaihua San on the general survival status,the appearance and histopathological changes of colon,related inflammatory cytokines,NF-κB signaling pathway and microangiogenesis of UC rats were observed,so as to reveal the specific mechanism of Huaihua San in the treatment of UC.In this study,clinical trials and animal experiments were conducted to explore the intervention and efficacy of Huaihua San on UC,and the specific mechanism of the efficacy was studied.Method:In the clinical trial,60 patients were selected to meet the diagnosis criteria of Western medicine for UC,which was diagnosed as large intestinal damp-heat syndrome by TCM.According to the principle of randomization and control,60 cases were divided into experimental group and control group.The experimental group was treated with Chinese herbal decoction Huaihua San,and the control group was treated with Mesalachin sustainedrelease granules.Patients were treated for 4 weeks in strict accordance with the requirements.The differences of subjects in gender,age,disease course and other aspects need to be statistically analyzed to confirm the comparability of the data.To observe the effect of Huaihua San on TCM symptom improvement,related curative effect and blood inflammatory indexes.Animal experiments using dextran sodium sulfate(DSS),copy the rats UC model,were randomly divided into blank group,model group,Huaihua San highdose group,medium-dose group,low-dose group,western medicine mesalazine group,give corresponding drug intervention respectively,rats were observed before and after medication general survival situation,colon gross appearance,histopathological changes and the related inflammatory cytokines(IL-1 beta,IL-10)test.The protein and mRNA expressions of NFκB p65,IκBα and IKKβwere detected.VEGF,VEGI and bFGF were detected by Elisa and CD34 labeled microvessel density was detected by immunohistochemical method.Results:1.The clinical study on Huaihua San in the treatment of UC patients with large intestinal damp-heat syndrome showed that:after treatment with Huaihua San,the improvement of other symptom scores and total symptom scores in the experimental group was better than that in the control group(P<0.05)except the number of stool.The two groups had similar therapeutic effects on the improvement of Mayo total score and Baron score(P>0.05).After treatment,the therapeutic effect of single main symptom and overall syndrome in the experimental group was better than that in the control group(P<0.05).In terms of clinical comprehensive efficacy,the two groups had similar effects(P>0.05).The experimental group was superior to the control group in the improvement of blood indexes CRP,IL-1β IL-10 and TNF-α(P<0.05),and the treatment effect of the two groups was similar in the improvement of blood indexes ESR and IL-6(P>0.05),and no adverse events occurred after the end of the trial.2.The experimental study on the intervention of Huaihua San in UC rat model induced by DSS showed that:(1)colon length,DAI score,histopathological score and CDMI score of rats:after intervention with high,medium and low doses of Huaihua San and western medicine Meisalachin sustained-release granules,groups of rats colon length all have different degrees of improvement,the Huaihua San highdose group improved the most obvious,was extremely significant difference compared with the western medicine group(P<0.01).In terms of DAI score,DAI score in high-dose and medium-dose Huaihua San groups was lower than that in western medicine group(P<0.05).In terms of histomathological score,the effect of high-dose and medium-dose Huaihua San group was similar to that of western medicine group,and the difference was not statistically significant(P>0.05).In terms of CDMI score,the high,medium and low dose groups of Huaihua San were similar to the western medicine group,and the difference was not statistically significant(P>0.05).(2)inflammatory cytokines:after drug intervention,compared with model group,Huaihua San group and western medicine group of IL-1β,IL-6,IL-10,the TNF-α level also have changed,among them,Huaihua San high-dose group reduce IL-1β,IL-6,TNF-α level better than western medicine group,in elevated inflammatory factor levels of IL-10 is superior to western medicine group(P<0.01).(3)IκBα,IKKβ,NF-κB p65:effects on gene expression:after drug intervention,compared with model group,Huaihua San group and western medicine group IκBα,IKKβ,NF-κB p65 gene expression has changed,and Huaihua San high-dose group and medium-dose group of rats IKKβ,NF-κB p65 gene expression was lower than western medicine group(P<0.01),IκBαgene expression was higher than western medicine group(P<0.01).Effect on protein expression:after drug intervention,compared with model group,the expression level of IκBα in Huaihua San high-dose group and western medicine group was increased(P<0.05),and the effect of the two groups was similar(P>0.05).The protein expressions of IKKβand NF-κB p65 in Huaihua San high-dose,medium-dose and western medicine groups were decreased(P<0.01).In IKKβ,NF-κB p65 protein expression levels,Huaihua San high-dose group with similar effect of western medicine group(P>0.05),(4)VEGF,VEGI,bFGF and microvasscular density counts:After drug treatment,compared with model group,serum VEGF in each group of Huaihua San and western medicine group decreased,and there was no significant difference in VEGF level between Huaihua San high-dose group and western medicine group(P>0.05).After drug treatment,serum VEGI increased,and there was no significant difference in VEGI level between Huaihua San high-dose group and western medicine group(P>0.05).After treatment,serum bFGF increased,and there was no significant difference in bFGF level between Huaihua San high-dose group and western medicine group(P>0.05).Immunohistochemical staining results showed that compared with the normal group,the MVD in the model group was increased,the difference was extremely significant(P<0.01);compared with the model group,the MVD in the each group of Huaihua San and western medicine group was decreased,the difference was extremely significant(P<0.01);the MVD in the Huaihua San high-dose group was lower than that in the western medicine group,the difference was significant(P<0.05).Conclusion:Huaihua San has obvious clinical efficacy in the treatment of UC patients with large intestinal damp-heat syndrome.It has been confirmed that Huaihua San,the representative prescription for the treatment of "intestinal wind bleeding ",can play the role of "clearing heat and cooling blood,draining the wind to down the qi,regulating qi and blood" and has the effect of treating UC,which is clinically effective and safe.Animal experiments showed that Huaihua San increased the expression of inhibitory protein IκBαand decreased the expression of IKKβ by inhibiting the activation of NF-κB pathway,thereby inhibiting the expression of pro-inflammatory factors IL-1β,IL-6 and TNF-α,and increasing the level of anti-inflammatory factor IL-10.From the perspective of increasing angiogenesis in inflammatory response,it was confirmed that Huaihua San can effectively reduce the expression of VEGF,increase the expression of VEGI,reduce microangiogenesis,increase the expression of bFGF,and promote mucosal repair and healing,which may be the specific mechanism of Huaihua San in the treatment of UC. |