Integrated Investigation On The Efficacy And Mechanism Of Sini Decoction Against Ulcerative Colitis

1 AimsThe classic famous TCM(traditional Chinese medicine)has always played an important role in the traditional Chinese medicine diagnosis and treatment system.The precise clinical application and positioning of classic famous TCM are conducive to the expression its clinical value.This article focus on the classic famous TCM Sini Decoction(SNT)as an example,and adopts the "network pharmacology prediction-animal experiment verification-proteomics analysis" integrated research strategy to target the main disease of XIALIQINGGU.This study provides the research and re-evaluation of indications,the role of SNT treat in ulcerative colitis(UC),drug efficacy evaluation and preliminary mechanism research,to lay the foundation for the clinical repositioning of SNT.2 Methods and results2.1 Prediction of the effect of Sini Decoction based on bioinformaticsThe chemical constituents of the SNT group of Chinese medicines were collected by the BATMAN-TCM database,and the "component-target" prediction was performed.The molecular,functional and biological pathways involved in the potential drug targets of SNT were significantly enriched by GO and KEGG,and the OMIM,TTD and DAVID 6.8 databases were used for multi-source significant enrichment analysis of SNT potential digestive diseases.According to the database,a total of 335 chemical components of SNT triglyceride were collected,and a total of 1009 corresponding targets were predicted,and there were synergistic targets between the chemical components.Among them,there were 143 common targets for Aconiti Lateralis Radix Praeparata and Zingiberis Rhizoma;146 for common targets between Aconiti Lateralis Radix Praeparata and Glycyrrhizae Radix et Rhizoma;396 for common targets between Zingiberis Rhizomaand Glycyrrhizae Radix et Rhizoma;and 110 for common target of three drugs,suggesting potential for three drugs in SNT.The concept of compatibility.Through functional enrichment analysis,it was found that SNT specifically acts on digestive organs such as liver,stomach,colon and small intestine.In addition,SNT is also enriched in the colorectal cancer cells,duodenum,gastric cancer cells,rectum,esophagus,intestine and other digestive systems.The results of clinical disease enrichment analysis involving SNT suggest that SNT affects digestive diseases involving UC.Finally,based on the previous ancient clinical medical case mining,Chinese medicine clinical drug experience analysis,combined with network pharmacology analysis results and the frequency of disease occurrence,taking into account the disease transformation law and the feasibility of subsequent experiments,through the integration analysis,prompted the potential association between the digestive system(especially UC)with the XIALIQINGGU clinical position of SNT.2.2 Sini Decoction in the study of TNBS-induced acute ulcerative colitis in ratsIn view of the above predicted results of clinical new positioning,in order to investigate the relationship between SNT and UC,the acute UC model induced by 2,4,6-trinitrobenzenesulfonic acid(TNBS)was selected.Conduct pharmacodynamic studies and validation.According to the body weight of the rats,the TNBS induction group and the blank group were randomly divided into groups,and the TNBS induction group was subjected to TNBS modeling.Rats with successful modeling were randomly divided into model group,SNT(19.2,9.6,4.8 g·kg-1)group and SASP(Sulfasalazine Enteric-Coated Tablets,SASP)group according to body weight and modeling severity.The administration was started three days after the modeling.The SNT group was administered intragastrically according to the respective doses.The positive drug SASP group was intragastrically administered with a SASP suspension at a dose of 0.5 g·kg-1,and the rest was given an equal volume of physiological saline.The state of UC rats was evaluated during the experiment using general observation and disease activity index(DAI).After the end of the administration,the serum levels of IL-6,TNF-a and PGE2 were detected by ELISA.The levels of NO in the serum of each group were detected by colorimetry.Colonic mucosa damage index(CMDI)evaluation and HE were performed.Staining was used to evaluate the results of colonic injury in rats.The results showed that compared with normal group,UC rats CMDI increased significantly,serum levels of TNF-?,PGE2 and IL-6 increased significantly,and the expression of NO was up-regulated,suggesting successful modeling.SNT can significantly improve diarrhea,blood in the stool,improve colonic mucosal congestion,edema,injury and adhesion in UC rats,and can significantly down-regulate the expression levels of serum TNF-a,PGE2 and IL-6,and have a tendency to down-regulate NO expression.In addition,compared with the positive drug SASP group,SNT groups have an advantage in down-regulating the expression levels of TNF-a,PGE2 and IL-6.It is preliminarily believed that SNT 19.2,9.6,and4.8g·kg-1 groups can significantly improve the symptoms of moderate to mild UC intestinal mucosal injury,and exert anti-UC effects by down-regulating the expression levels of TNF-?,PGE2,IL-6 and NO.And the efficiency is better than SASP.Based on this,this paper verifies the pharmacodynamic effect of SNT intervention in UC from the perspective of animal experiments,and further provides experimental basis for the relationship between the digestive system(especially UC)with the XIALIQINGGU clinical position of SNT.2.3 Sini decoction intervention mechanism and key target of ulcerative colitisTo further analyze the mechanism of SNT intervention in UC,the effect of SNT was tested and confirmed,and proteomics was used to analyze the expression of distal colon protein in rats in the blank group,model group and SNT group(9.6 g·kg-1).In the case,differential protein screening was performed after multiple test calibration,and GO and KEGG functional enrichment analysis was combined with literature research to screen potential key proteins and validate.After analysis,78 SNT-mediated key differential proteins of UC,including 26 up-regulated proteins and 52 down-regulated proteins,were screened to explain the multi-target regulation of SNT on UC.Combined with bioinformatics analysis,it is preliminarily believed that SNT may intervene by interacting with protein interaction networks in the intima,extracellular regions,and cytoplasmic vesicles.SNT-regulated proteins are involved in glycosaminoglycan binding,sulfur compound binding,calcium ion binding,TLR4 binding,extracellular structural organization,exocytosis,extracellular matrix tissue,neutrophil degranulation,and cell secretion.A total of 16 key proteins such as CRP,MPO,CAMP,S100A9,S100A8,SPARC,COL12A1 and ELANE were screened out.CRP and COL12A1 are focused as potential targets to be screened,with CRP being a known clinically UC biomarker.Through the ELISA experiment,the down-regulation effect of SNT on the commonly used biomarker CRP was confirmed,and the down-regulation effect of SNT on the new potential key target COL12A1 was verified.It is verified that SNT may play a role in interfering with UC by reducing protein expression in colon and serum CRP and COL12A1,and that COL12A1 has potential as a new potential target.In summary,this study further confirms the association betweenthe digestive system(especially UC)with the XIALIQINGGU clinical position of SNT.,and preliminarily clarifies the mechanism of SNT intervention in UC.3 ConclusionIn this study,the integration strategy was used to initially position the digestive system disease UC as theclinicaldiseaseofSNT XIALIQINGGU.Besides,the therapeutic efficiency was observed.It was found that SNT can recover colonic mucosal injury,improve stool viscosity,inhibit blood in the stool,and down-regulate TNF-a,PGE2,and IL-6 expression in UC model ratsserum,better than SASP.In addition,SNT also exerts a pharmacological effect on UC by regulating 78 vital differential proteins of UC,and CRP and COL12A1 can be used as potential vitaltargets for SNT treatingfor UC.The integrated research strategy has clinically repositioned SNT and provided an effective demonstration of the precise clinical application and the position of the classic famous TCM.