| Objective: The aim of this study was first to describe the clinical efficacy of CYT in combination with mesalazine compared with mesalazine,then to analyze the molecular mechanisms and targets of CYT in the treatment of ulcerative colitis(UC)in the context of modern cyberpharmacology and molecular docking,and then to validate the possible mechanisms of CYT in the treatment of UC mice and whether CYT exerts a protective effect on the intestinal mucosa of the colon in mice by means of animal experiments.Methods: Patients diagnosed with ulcerative colitis(mild to moderate)in the active stage and classified as large intestine dampness-heat syndrome were collected from the outpatient department and treatment area of Hepato-spleen Gastroenterology Department of the First Clinical Hospital of Jilin Academy of Traditional Chinese Medicine from September 2022 to October 2023.During treatment,TCM symptom score,Mayo score,TCM symptom efficacy score,total clinical efficacy score,and IBDQ score of patients before and after treatment were collected,and the results were statistically analyzed to evaluate the clinical efficacy of intestinal therapy decoction.By using network pharmacology and molecular docking methods,the active ingredients of UC and their related gene targets were collected from the relevant network database,and the "drug active ingredients-target network map" was constructed to explore the possible mechanism of UC treatment.Collect intersections of UC disease and drugs;Collect information on the intersection of UC disease and medication.Cross targets of UC diseases and drugs were collected and Wayne’s map was drawn.Then the cross-targets were imported into the corresponding database for protein interaction network analysis(PPI,GO,KEGG).The core targets,signaling pathways,biological processes(BP),cell components(CC),and molecular functions(MF)of Changy decoction were screened.The results were visually analyzed by molecular docking method UC mouse model was established in animal experiments,and2.5% dextran sulfate sodium(DSS)was used to induce modeling for 7 days.After the modeling with DSS was finished,the UC model mice were treated with high and low doses of Intestinal Healing Soup for 10 days,and the body weights,colonic lengths,and disease activity indices(DAIs)of the mice were compared before and after the treatments.The serum and colon tissues of mice were collected,and hematoxylin-eosin staining(H&E staining)was used to observe the damage of the colon tissues;IHC immunohistochemistry was used to observe the changes in the expression of ZO-1,Occludin,and MUC2 in the colon tissues of mice.The serum levels of interferon γ(TNF-α),interleukin 6(IL-6),interleukin 1β(IL-1β),interleukin 8(IL-8),and interferon γ(INF-γ)were detected by Elisa,and the mRNA expressions of IL-8,INF-γ,TNF-α,IL-6,and IL-1β were detected by realtime quantitative PCR;Western-bolt method was used to detect AKT,p-AKT,IκB-α,p65,pp65 protein contents in colon tissues;Results: A total of 64 cases were collected in the clinical observation of Intestinal Healing Soup in the treatment of ulcerative colitis,and the results showed that the Chinese medicine clinical symptoms of the patients,including abdominal pain,defecation,abdominal discomfort,and mucus,pus,and blood stools,were significantly improved,and the Mayo score of the experimental group of patients was significantly reduced after treatment;the points of the Chinese medicine major and minor symptoms of the experimental group of patients were significantly reduced after treatment;the Chinese medicine symptomatic effects were significantly better than that of the control group;the IBDQ scores showed significant improvement after treatment,and all of them were statistically different;the total clinical effective rate of the experimental group in the treatment of UC patients reached 96% than that of the control group.The results of the IBDQ score showed that the experimental group improved significantly after treatment,and all of them were statistically different;the total clinical effectiveness rate of the experimental group for the treatment of UC patients reached 96%,which was better than that of the control group.The network pharmacology study showed that 239 active ingredients were obtained after searching,including 9 Baihe,2 Paeonia lactiflora,66 Salvia miltiorrhiza,13 Dijinzhao,13 Diyu,15 Poria,92 Glycyrrhiza glabra,14 Huanglian,6Sophora,and 9 Coix lacryma.After combining the targets obtained from the TCMSP database and removing the duplicates,a total of 246 targets were obtained from the CYT compound,and a total of 127 targets were obtained after intersecting with the disease targets,and 10 important active ingredients and 5 important targets were obtained by screening according to the ranking of Degree value.The results of the GO analysis showed that intestinal healing broth was involved in the activation of chemoreceptors,cellular senescence,platinum resistance,etc.,respectively(BP),affecting the membrane,and influencing the biological processes(BP),and affecting the membrane.The results of KEGG analysis showed that Gutta-percha was involved in the regulation of several signaling pathways including PI3K-Akt,NF-κB,AGE-RAGE,and TNF,but mainly related to PI3K-Akt,NF-κB,AGE-RAGE,and TNF,according to its P-value.based on its P value,it was found to be mainly associated with the PI3K-Akt signaling pathway.The molecular docking results showed that the top five core targets of the screening had good binding activities with their corresponding active ingredients.The results of animal experiments showed that the Intestinal Healing Soup could effectively reduce the disease activity score of UC model mice established by DSS,reduce the weight loss of UC mice,and alleviate the shortening of the colon in UC mice.In the results of H&E staining and immunohistochemistry,it was observed that Intestinal Healing Soup could effectively improve the intestinal inflammatory infiltration in UC model mice,promote the healing of colonic mucosal ulcers,reduce the edema of the colonic intestinal tract,and effectively up-regulate the expression of three tight junction proteins,namely,ZO-1,Occludin,and MUC2,in UC mouse colon,and Intestinal Healing Soup could also make the colonic tissues in mice TNF-α,IL-6,Western Blot results showed that Gutta-percha had a certain inhibitory effect on the PI3K-Akt/NF-κB pathway.Conclusion: In the process of clinical treatment of ulcerative colitis,the test group can significantly reduce the patient’s Mayo score,improve the IBDQ score,and improve the patient’s quality of life;the clinical symptom score of Chinese medicine in the test group is also significantly due to the patients in the control group,and the total effective rate of the efficacy of the test group reaches 96% after the statistics.And there were no adverse reactions or side effects in the whole treatment of this trial,the effect was good.The results of network pharmacology and molecular docking showed that many active ingredients in Intestinal Healing Soup for UC may be the key to treating UC.After GO and KEGG analyses,it was found that Gutta-percha soup might be involved in the occurrence and progression of UC through cellular component(CC),biological process(BP),and molecular function(MF)processes,and Gutta-percha soup might be mainly involved in the regulation of the PI3K-Akt/NF-κB pathway to play a therapeutic role in the treatment of UC.The molecular docking results showed that the core target of Gutta-percha soup has a good affinity with the active ingredient of the drug.Finally,the up-regulation of the expression of ZO-1,Occludin,and MUC2 in the colon tissues of UC mice confirmed that the Intestinal Healing Soup could protect the colon mucosa of UC mice,reduce the damage of the intestinal mucosa,and improve the repair of colon tissues to a certain extent.In the present experiment,it was shown that Gut Guai Tang could effectively inhibit the phosphorylation of Akt/p-65 in the PI3K-Akt/NF-κB signaling pathway,and reduce the release of the related inflammatory factors,TNF-α,IL-6,IL-1β,IL-8,and INF-γ,which could reduce the intestinal inflammatory response and promote the healing of intestinal mucosa.Clinical and animal experiments have verified that intestinal healing soup has a certain therapeutic effect on UC,which is worthy of clinical promotion and application. |
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