| Objective: In this paper,the pharmacodynamic material basis and therapeutic mechanism of BXD for the prevention and treatment of inflammatory colon cancer were investigated using the classical prescription BXD,with a view to elucidating the pharmacodynamic effect,effector substances,and mechanism of action of BXD for the treatment of inflammatory colon cancer through various techniques and methods,such as pharmacodynamics,serum medicinal chemistry,cyber-pharmacology,serum non-targeted metabolomics,proteomics,and intestinal microbiology,and so forth,so as to evaluate the effectiveness and safety of BXD for the treatment of inflammatory colon cancer and to provide a research basis for the clinical application and further development of BXD.The aim is to elucidate the efficacy,effector substances and mechanism of action of BXD in the treatment of inflammatory colon cancer,in order to evaluate the effectiveness and safety of BXD in the treatment of inflammatory colon cancer,and to provide a research basis for the clinical application and further development of BXD.Methods: 1.A mouse model of colonic inflammatory colon cancer(CAC)was constructed using oxidized azomethine methane(AOM)/dextran sulfate sodium(DSS),mice were observed for general condition,organ indexes,and mouse tissues were pathologically analyzed using H&E staining and immunofluorescence.2.Liquid-liquid mass spectrometry(LLM)methods were used for systematic qualitative analysis of the BXD and drug-containing sera,and the second level mass spectra were performed according to the positive-negative ion mode Fragmentation ion information and retention time,the source and information of the components were determined by self-constructed library and consulting related literature and database search.Combining the literature reports and mass spectrometry data,we used databases such as the Chinese medicine composition database,published literature,and the website of Gene cards,to collect the active ingredients,targets of action,and obtain genes related to colitis colorectal carcinoma of the various medicines in BXD.The potential targets of BXD for the treatment of colitis colorectal cancer were analyzed by PPI network construction,bioprocess enrichment analysis(BP)and KEGG pathway enrichment analysis,and the regulatory network of "BXD-constituents-targets-disease" was constructed.3.AOM/DSS was used to construct the CAC model of mice,and the contents of the cecum were taken for the extraction of total intestinal microbial DNA and the extraction of total intestinal microbial DNA.The total DNA of intestinal microorganisms was extracted and detected,PCR amplification of bacterial 16 S r RNA gene was performed,sequencing was performed,and Alpha and Beta diversity analyses were performed on the measured data to obtain the differential flora of the intestinal flora regulated by BXD.Blood was extracted from the eyeballs of the mice with CAC,and the liquid quality was detected.metabolite screening,differential metabolite correlation analysis,and performing intergroup variability analysis to obtain differential biomarkers and metabolic pathways in BXD-regulated CAC mice.The colon tissues of CAC mice were collected and subjected to liquid-mass spectrometry for protein qualitative and quantitative analysis,biochemical analysis,statistical analysis,comparison of differential proteins,and PCR verification of key differential proteins.Results: Pharmacodynamic studies have shown that BXD can inhibit inflammatory response and tumor development in CAC model mice without hepatotoxicity and nephrotoxicity,and can regulate the expression of CAC prognostic-related factor HIF-1α,and affect the expression of serum inflammatory factors TNF-α,IFN-γ,IL-6,IL-1β,and the inflammation-related pathways TNF-α and IFN-γ,and thus exert anti-CAC effect.A total of 289 chemical components,including 81 flavonoids,35 alkaloids,35 phenylpropanoids,35 terpenoids,and 103 other compounds,were detected and identified by using liquid-mass spectrometry technology and serum medicinal chemistry and network pharmacology,of which 142 compounds were blood-entry prototypical components,and 8compounds were metabolites of prototypical components.Four key genes,CD38,CXCL8,HIF-1 α and PTGS2,were analyzed in the CAC prognostic model,and four key genes regulating the prognosis of CAC were analyzed in the BXD-chemical component-target-disease regulatory network,and molecular docking validation was performed to determine that aloe barbadensis,apigenin,sennosides,geraniol,wild scutellarin were the key genes regulating CAC prognosis.lignans,and wild baicalein as the potential pharmacological material basis for the prevention and treatment of colitis colon cancer by hemixia laxative heart soup.The results of intestinal microbial assay showed that half-sia laxative heart soup could regulate Firmicutes＿D,Bacteroidota,Firmicutes＿A,Actinobacteriota,Desulfobacterota＿I,Proteobacteria,Deferribacterota,Patescibacteria,Verrucomicrobiota,Campylobacterota,and microbial abundance at the phylum level,and can regulate Muribaculaceae,Erysipelotrichaceae,Lachnospiraceae,Lactobacillaceae,Desulfovibrionaceae and other microbial abundance at the level of genera.The biopathway enrichment analysis revealed that Hanxia Laxin Tang could regulate various biopathways such as biosynthesis,assimilation process,detoxification,glycoprotein pathway and other biopathways,and further analysis revealed that Burkholderiaceae A,Further analysis showed that Burkholderiaceae A,Parasutterella,Burkholderiales,Oscillospirales and Ruminococcaceae may be the key flora for the therapeutic effect of Hanxia Laxative Heart Tang.Metabolomic analysis yielded that all serum endogenous metabolite levels in mice tended to be regressed after the intervention of hemixia laxative heart soup,and the main differential metabolites were Adenosine,Lyso PE(16:1(9Z)/0:0),N-Cyclohexyl-N’-(4-pyridinylmethyl)thiourea,18-alpha-Glycyrrhetinic acid,Coptisine cation,etc.Pathway enrichment analysis revealed that the differential metabolites were mainly enriched in ABC transporters,Alcoholism,Arginine and proline metabolism,Glycine,serine and Further metabolite-gut microbial association analysis revealed that Hanxia Laxing Xin Tang could play a therapeutic role in the treatment of inflammatory colon cancer through metabolite regulation,gut microbial regulation and other pathways.Proteomics study showed that there were 383 up-regulated proteins and 237down-regulated proteins in the colon tissue proteins of model mice,and after the intervention of Hanxia Laxin Tang,there were 30 different proteins up-regulated and 82 different proteins down-regulated,and the pre-cancerous administration of Hanxia Laxin Tang also showed a certain regulatory effect,suggesting that the Hanxia Laxin Tang has the potential effect of preventing tissue cancer,and the results of the GO functional enrichment showed that the Hanxia Laxin Tang can GO functional enrichment results showed that Bianxia Laxing Heart Tang could play preventive and therapeutic roles by affecting the regulation of biological process,protein binding,organic cyclic compound binding and other functions in CAC model mice,and the KEGG pathway enrichment showed that its main regulatory pathways include mmu04610 Complement and coagulation cascades,mmu00980 Metabolism of xenobiotics by cytochrome P450,mmu05171 Coronavirus disease-COVID-19,etc.q PCR validation results showed that Half-summer diarrhea heart soup can affect the expression of complement and coagulation cascade reaction signaling pathway by regulating the expression levels of four proteins,C3,Cfh,Cpb2 and Fgg,further indicating that BXD may play a role in preventing colitis colon cancer by regulating multiple targets and pathways.Conclusion: The results of pharmacodynamic study showed that the soup could prevent CAC,regulate the expression of serum inflammatory factors TNF-α,IFN-γ,IL-6,IL-1 β and inflammation-related pathways TNF-α and IFN-α,and interfere with the expression of prognostic factors in colon inflammatory colon cancer,and inhibit the inflammatory response and tumor development in model mice,without hepatotoxicity and nephrotoxicity.The potential pharmacodynamic substances are aloe emodin,apigenin,shannelin,geraniin and baicalein.Mechanism of action studies showed that,Pinellia heart soup can regulate the abundance of microorganisms such as Muribaculaceae,Erysipelotrichaceae,Lachnospiraceae,Lactobacillaceae and Desulfovibrionaceae,And then play a therapeutic role;Regulating the serum metabolites levels of mouse Adenosine,Lyso PE(16:1(9Z)/ 0:0),N-Cyclohexyl-N’-(4-pyridinylmethyl)thiourea,18-alpha-Glycyrrhetinic acid,And then play a therapeutic role;Metabolite-gut microbial association analysis found that,Changes in gut microbial levels can affect metabolite changes.Pinellia pinellia diarrhea heart soup can adjust the expression level of tissue proteins and has a potential effect in preventing tissue carcinogenesis.The potential targets against colon inflammatory colon cancer are C3,Cfh,Cpb 2,Fgg,HIF-1 α,NF-κ B,IFN-γ,etc.In conclusion,pinellia diarrhea heart soup can play a role in treating colon inflammatory colon cancer by affecting intestinal microbial levels,serum metabolites levels and protein expression levels.The potential pharmacodynamic substances are aloe emodin,apigenin,sananin,curcumin,and the potential targets are C3,Cfh,Cpb 2,Fgg,HIF-1 α,NF-κ B,IFN-γ,etc. |
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