Efficacy And Safety Of Mesalazine And Sulfasalazine For Treating Ulcerative Colitis:A Meta-Analysis

Objective:It is reportedly evidenced that the etiology and diseasing mechanism of ulcerative colitis(UC)have not been fully elucidated,and the disease has become one of the clinically refractory chronic diseases.Clinically,sulfasalazine and mesalazine were commonly-used drugs for clinical treatment of UC.However,there are still some controversies to the clinical efficacy and safety of these anti-UC medicines.To systematically compare the therapeutic effects of these drugs,this chapter was designed to apply the Meta-analysis methodology via assessing the currently completed clinical randomized trials.We tried to identify the drug efficacy and safety by using evidence-based medicine analysis,and then to provide novel ideas for clinical anti-UC medication guidance and optimization of combined medication.Materials and methods:1.Firstly,we established inclusion and exclusion criteria according to requirement and PICOS principle.The search strategy was to screen subject words combined with free words in Chinese and English databases.Then,the researcher filters references retrieved from the databases.Non-conforming and duplicated documents should be deleted.Finally,the quality of the included studies was determined by evaluator and the data was extracted using a specially designed data extraction table,which included writer’s name,sample size,therapy sessions,patient baseline characteristics.2.Methodological quality evaluation of the included studies was carried out using the Cochrane System Evaluation Manual.The risk of bias for each study was assessed against the criteria.The Review Manager 5.3 software was used for data analysis and graphics.Forest plot of meta-analysis could be employed to compare the efficacy and safety of those two-drugs and funnel plot could be applied to evaluate publication bias.Results:1.A total of 26 literatures/references were included in present meta-analysis,all of which randomized controlled trial.The total sample sizes comprised 4012 cases,in which there were 2057 cases in mesalazine group and 1955 cases in sulfasalazine control group.2.The meta-analysis results demonstrated that total-effective rate of mesalazine group was more significant than that in control group,and the difference was statistically significant.The adverse-effect rate of mesalazine treatment group was significantly lower than that in sulfasalazine group,featured with statistically significant differences.Conclusions:1.Mesalazine contributed to relatively better therapeutic action than sulfasalazine treatment for anti-UC.2.For assessment of treatment safety,the adverse-reactions of mesalazine for anti-UC were less than sulfasalazine intervention.Objective:In order to provide preclinical experimental data,this chapter intends to build up an in vivo model of DSS-induced UC mice,and then to evaluate and compare the pharmacological activities of mesalazine and sulfasalazine in the treatment of UC via basic pharmacological experiments.16 S r RNA sequencing technology was applied to detect and analyze the abundance and composition of intestinal flora in each group,as well to compare the damaged intestinal flora functions of mesalazine-and sulfasalazine-treated mice.We aimed to innovatively evaluate the pharmacological activities of mesalazine and sulfasalazine from the perspective of the microenvironment in intestinal flora,and then to provide a theoretical basis for joint supplement of beneficial bacteria for clinical anti-UC actions.Materials and methods:1.C57BL/6J mice were randomly divided into 6 groups(n=10)after building up the DSS-induced UC mice.The mesalazine group and DSS+ mesalazine group were intragastrically given with 0.75 g/kg per day,sulfasalazine group and DSS+ sulfasalazine group were treated via lavage with 0.6 g/kg per day.The body weight of mice was recorded every day and were sampled after 7 days.2.Weight changes,colon length,bloody stool and HE staining scores of colon were observed.ELISA determination for inflammation cytokines in colon tissue homogenate were conducted.The levels of serum ALT,AST were determined,liver and kidney tissues were checked through HE stains.3.Immunohistochemical and immunofluorescence stains and analysis were used for assessment of the expressions of Bax,Bcl-2,IκBα,NF-κB p65 in colon tissue.4.Whole metagenomics(16S r RNA sequencing)was employed to profile the composition of gut microbiota in mice.The anti-UC actions of mesalazine and sulfasalazine treatments were further calculated and compared through the changes of colon microbiota.Results:1.After successfully establishing UC model,the body mass of UC mice gradually decreased,and blood in the stool could be observed visibly.Mesalazine could effectively reverse the weight loss caused by DSS,instead,sulfasalazine resulted in no obvious effect on weight loss in UC mice.2.The autopsy displayed that the liver and kidney enlargements in UC mice.Serum ALT and AST levels were significantly increased,indicating that mesalazine and sulfasalazine treatments were found with certain toxic effects on liver and kidney.Notably,the liver or kidney damage caused by sulfasalazine was more serious than that in mesalazine.3.Immunohistochemistry and immunofluorescence staining experiments mutually corroborated the pharmacological activities of mesalazine and sulfasalazine against UC.The treatments coul reduce the positive expressions of endogenous Bax and NF-κB p65 proteins in colon tissues,and significantly increase the positive expressions of Bcl-2 and IκBα.Significantly,the pharmacological actions of mesalazine were better than sulfasalazine treatment.4.The findings/data of 16 S r RNA sequencing analysis showed that the abundance and uniformity of the intestinal microbial community increased following the treatments.After mesalazine treatment,the levels of Enterobacter and Laurespirillum were increased,however,the improvement of sulfasalazine resulted in no statistical significance.Conclusions:1.Mesalazine and sulfasalazine treatments had some toxicities to liver and kidney organs,however,the side-effects induced by mesalazine were less than those in sulfasalazine intervention.2.DSS-induced UC was related to the abnormal expressions of Bax and Bcl-2(apoptosis),NF-κB p65 and IκBα(inflammatory stress).Mesalazine and sulfasalazine could regulate the expressions of these effectors to exert anti-UC pharmacological activities,and the pharmacological benefits of mesalazine was better than these in sulfasalazine treatment.3.Mesalazine and sulfasalazine could suppress the decrease of intestinal microbial abundance in UC mice.In addition,mesalamine could also reconstruct the abundance of other beneficial bacteria.Mesalazine exerted anti-UC pharmacological actions via increasing beneficial bacteria to ameliorate the energy utilization and intestinal microenvironment.Objectives:In our present report,we first applied network pharmacology-derived bioinformatics method to systematically predict the core targets,biological processes and molecular mechanisms of mesalazine for treating UC.Moreover,the computer molecular docking method was applied to verify and clarify the anti-UC pharmacological targets of mesalazine.Meanwhile,the expression activities of these anti-UC pharmacological targets were experimentally verified using UC mouse model.We aimed to uncover the pharmacological targets and mechanisms of mesalazine-anti-UC action,and to provide a new theoretical basis for mesalazine-anti-UC.Materials and methods:1.All targets/genes of mesalazine and UC were harvested via the network databases of drug and disease prior to intersection targets being screened out and identified.All core targets were harvested by using corresponding software and included algorithms,and mesalazine-UC interaction network diagram was plotted respectively.2.Gene ontology(GO)annotation and KEGG assay with current core targets were determined for visualizing the "drug-target-GO-KEGG-disease" network relationship to clarify the bioinformatics characteristics of mesalazine-anti-UC.Structurally molecular docking detection was conducted to analyze the binding activity/capability of mesalazine and core target protein sites in UC.3.We successfully established DSS-induced UC mouse model to test the anatomical features,ALT kit,ELISA determination and HE stain.The results of network pharmacology and molecular docking were identified with immunostaining and immunoblotting assays.Results:1.A total of 175 therapeutic targets and 2711 UC-diseased targets were well-identified.75 shared targets of mesalazinne for treatment of colitis were obtained and showed in Venn diagram.According to exclusion criteria and algorithm,total 9 core targets were screened out and identified,comprising ALB,TNF,INS,PTGS2,MMP9,IL1 B,EGFR,HSP90AA1,MMP2.2.By use of GO and KEGG assays,we concluded that effectively altered biological processes and signal pathways exerted by mesalazine for treating UC.The bioinformatic findings of mesalazine-anti-UC were principally implicated in the modulation of inflammation and immune response,antigen processing and presentation,MAPK signal pathway and IL signal pathway.More notably,structural biology analysis highlighted the effective binding capacity of mesalazine to different functional sites in TNF,PTGS2,IL1 B,and EGFR core proteins.3.We observed the weight loss,blood in the stool,and shortened colon length in UC mice induced by DSS.Moreover,DSS-induced UC mice caused certain degree of liver or kidney damage.Routine HE staining demonstrated inflammation infiltration and cell necrosis in the intestinal tissues.4.The biochemical testing results manifested that the contents of IL-1α,IL6,IL8 and TNF-α in serum and colon homogenate samples in UC mice were significantly greater than those in normal mice.Specially,low-dose mesalazine treatment could effectively reduce the content of inflammatory factors in UC mice,while the improvement effect of high-dose mesalazine group showed no statistical significance.5.In order to experimentally verify the findings of bioinformatics analysis,the endogenous intestinal TNF-α,PTGS2,IL1 B and EGFR protein expressions in mesalazine treatment were significantly reduced when compared with these in UC mice.Conclusions:1.The bioinformatics findings from network pharmacology and molecular docking were employed to predict and clarify the core and pharmacological targets of mesalazine for treating UC,and the pharmacological actions and mechanisms associated with biological processes and signaling pathways that regulating immune function and inflammatory stress were uncovered respectively.2.Experimental data indicated that low-dose mesalazine treatment exerted better efficacy than that in high-dose,and underlying pharmacological actions are linked to the functional regulation of TNF-α,PTGS2,IL1 B,and EGFR protein activities.Objective:In our current report,an integrated omics strategy including transcriptomics and metabolomics was applied to uncover the therapeutic targets and molecular regulating mechanisms of mesalazine for treating UC on the basis of comprehensive analysis of high-throughput data.We further verified the omics analysis results/discovery through in vivo experiments.This chapter aimed to clearly reveal new/potential targets of mesalazine for treating UC,and then to lay a scientific theoretical foundation for the development of anti-UC research and development in combination with new target-based agents(such as chemokine inhibitors and T cell activators).Materials and methods:1.In transcriptomics analysis,we collected the colorectal tissues of each group with mice,and then extracted total RNAs and built a c DNA library.After transcriptome being sequenced,we analyzed and collected differentially expressed genes by use of algorithm with log2(Fold change)>1,P<0.05 to identify the significantly different genes.2.In metabolomics analysis,we first applied organic reagents to extract intestinal tissue metabolites after protein precipitation,and then performed sample separation and detection using liquid mass spectrometry(LC-MS).The mass spectrometry data and information were processed via meta X,XCMS software,and the substance qualitative and biological function annotations were carried out through HMDB,KEGG and in-house databases.The screening conditions were designed with q<0.05 and VIP≥1,and the output of corresponding charts in difference metabolites were compared and analyzed.3.Further,the main differentially expressed genes of transcriptomics analysis have been experimentally verified,including quantitative protein expression analysis(such as Western blot,ELISA).Results:1.Compared with the UC model group,mesalazine treatment resulted in a total of 1663 differentially expressed genes,including 262 up-regulated genes and 1401 down-regulated genes.GO and KEGG enrichment analyses indicated that the protective actions of mesalazine for treating UC were related to functional regulation of immune inflammatory response(such as regulating T cells,white blood cells and cytokine receptor pathways,etc.).In addition,IPA gene network mechanism assay further disclosed the inhibitory action of mesalazine on C-C motif chemokine ligands(CCL11 and CCL21)and C-X-C motif chemokine ligands(CXCL3 and CXCR2).2.Metabolomics assay displayed intestinal metabolite expression differences between the groups of UC model and mesalazine treatment,a total of 16 differential metabolites were screened and identified,including 9 up-regulated metabolites and 7 down-regulated metabolites.KEGG enrichment assay suggested that altered metabolic pathways exerted by mesalazine mainly included the unsaturated fatty acid pathway,the tryptophan metabolic pathway,and the serotonin metabolic pathway.3.By using ELISA assay,the intestinal expression of G-CSF in mesalazine treatment was significantly lower than that in UC group,and the expression of CXCL9 was greater than that in UC group.For semiquantitative analysis,the expressions of intestinal AOC3,MAP3K8,Ep300 following mesalazine treatment were down-regulated and the expressions of Beta catenin,Nox1,IL12RB2 were up-regulated.Conclusions:1.Mesalazine effectively ameliorated the immune function of UC mice,and regulated the expressions of CCL11,CCL21,CXCL3 and CXCR2 to suppress the progression of DSS-induced intestinal inflammation.2.At the metabolite level,mesalazine effectively ameliorated the metabolism of unsaturated fatty acids in the intestines,the metabolism of tryptophan,and serotonin,thereby improving the body’s metabolic function and exerting anti-UC pharmacological effects.3.Mesalazine can markedly exert potent roles by inhibiting or reducing G-CSF,AOC3,MAP3K8,Ep300 production and promting the expression of CXCL9,Beta catenin,Nox1,IL12RB2 for anti-UC.Additionally,experimental verification of transcriptomics analysis results/findings was completed in vivo.