Meta-analysis And Mechanism Study On Huangqin Tang In The Treatment Of Ulcerative Colitis

Objective:1.To systematically evaluate the efficacy and safety of HQT combined with mesalazine in the treatment of Ulcerative Colitis(UC).2.To explore the material basis of HQT for the treatment of ulcerative colitis(UC)and its possible mechanism of action based on the network pharmacology approach.3.To observe the intervention effect of HQT on DSS-induced ulcerative colitis(UC)mice and to investigate the effect of HQT on NLRP3/Caspase-1 signaling pathway in UC mice in order to elucidate its mechanism of action on UC.To provide scientific basis for the new drug development and clinical application of Huangqin Tang.Methods:1.The Chinese and English literature of randomized controlled trials(RCTs)of UC with HQT were searched using China Knowledge Network,Wanfang Data,Vipshop,PubMed,SinoMed,Embase,and Cochrane Library databases,and the search time was from the establishment of the database to January 2021.The included literature was evaluated for data extraction and risk of bias,and the efficacy and safety were evaluated using RevMan 5.3 software,and the quality of evidence was evaluated using GRADE.2.The potential active ingredients of HQT and their targets were obtained from TCMSP database,and the targets related to UC diseases were retrieved from Genecard and OMIM databases,and 93 potential targets of HQT for UC treatment were obtained by intersecting the two results,and the potential targets and active ingredients were imported into Cytoscape software to draw "HQT-active ingredient-potential target" network map.The potential targets and active ingredients were imported into Cytoscape software,and the network diagram of "HQT-active ingredient-potential target" was drawn to screen the key ingredients.Meanwhile,the potential targets were imported into STRING database,PPI network was constructed,and the core targets of HQT for UC treatment were screened.We used the Metascape platform to perform GO(gene ontology)biological process enrichment analysis and KEGG(KEGG pathway analysis)pathway annotation analysis on the intersection of drug and disease targets.3.C57BL/6J mice were randomly divided into blank group,model control group(DSS group),HQT-low,medium and high dose group(HQT-L group,HQT-M group,HQT-H group)and western medicine mesalazine group(western medicine group).The UC mouse model was established by freely drinking the DSS solution configured at 2.5%,and the HQT-L,HQT-M,HQT-H and western.drug groups were given low,medium and high doses of HQT and mesalazine suspension by gavage.An equal volume of distilled water was given to the blank group and DSS group by gavage.After 10 days of administration,the DAI score,colon length and colon histopathology were measured in each group,the serum IL-6,IL10,IL-1β and TNF-α contents were determined by ELISA,the expression of NLRP3 and Caspase-1 in colon tissue was determined by Q-PCR,and the expression of IL-6,IL-1β,NLRP3 and Caspase-1 in colon tissue was detected by immunohistochemistry.Results:1.A total of 6 studies with a total of 565 subjects were included,and Meta-analysis showed that HQT combined with mesalazine in treating UC significantly improved the cure rate[RR=1.56,95%Cl(1.23,1.98),P=0.0003]and overall effective rate[RR=1.24,95%CI(1.14,1.35),P=0.00001],which significantly reduced clinical symptom scores,but all with high heterogeneity.HQT combined with mesalazine modulated patients’ serum IL-6[SMD=-2.25,95%CI(-2.61,-1.88),P=0.00001],IL-10[SMD=2.01,95%CI(1.73,2.30),P =0.00001],IgA[SMD=-0.65,95%CI(0.96,-0.35),P=0.0001],IgG[SMD=-3.20,95%CI(-3.65,-2.75),P=0.00001]expression levels.HQT combined with mesalazine in the treatment of UC had a tendency to reduce adverse effects,but there was no statistical difference[RR=0.25,95%CI(0.06,1.10),P=0.07].All GRADE ratings of quality of evidence were low quality.2.The main active ingredients of HQT for UC treatment are quercetin,kaempferol,baicalein,etc.The core targets are AKT1,JUN,IL6,VEGFA,STAT3,MYC,CASP3,EGFR,etc.The treatment of UC with HQT mainly acts on biological processes such as cancer pathway,MAPK signaling pathway,TNF signaling pathway,insulin resistance,JAK-STAT pathway,Th17 cell differentiation,NF-kB and other signaling pathways,involving biological processes such as endotoxin response,regulation of apoptosis signaling pathway,regulation of small molecule metabolic process,reactive oxygen metabolic process,negative regulation of cell proliferation.3.The mice in the DSS model group showed significantly higher DAI scores and intestinal histopathology scores than normal mice,as well as significantly lower body weight,and intestinal pathology showed:multiple ulcer formation,irregular arrangement of glands around the ulcer,degenerative necrosis of columnar epithelial cells,reduction of cupular cells,and accumulation of deep chronic inflammatory cells in the lamina propria.Compared with the model group,the DAI scores of HQT groups and western medicine groups were significantly improved,among which the improvement was more obvious in HQT-M group,H group and western medicine group,and there was a statistical difference compared with the model group(P<0.05);the intestinal histopathology scores of HQT groups and western medicine groups were significantly decreased,among which the improvement was more obvious in HQT-M group,H group and western medicine group,and there was a statistical difference compared with the model group(P<0.05).In addition,compared with the blank group,the expression of NLRP3 and Caspase-1 in the colon tissue of the DSS model group was significantly increased,while the serum levels of IL-6,IL-1β and TNF-αwere significantly increased and IL10 was significantly decreased in the mice.Compared with the mice in the DSS model group,the expression of NLRP3 and Caspase-1 in the colonic tissues of the HQT groups and the western medicine group were significantly lower compared with the model group;while the serum levels of IL-6,IL-1β and TNF-α were significantly lower and IL10 was significantly higher in the HQT groups and the western medicine group,and the improvement was most obvious in the HQT-H group and the western medicine group,with statistically significant differences(P<0.01).Conclusions:1.HQT combined with mesalazine in the treatment of UC significantly improved the cure rate and overall efficiency,reduced intestinal Mammation and protect intestinal mucosa,and had the advantages of integrated traditional Chinese and western medicine.2.This study initially revealed the multi-component,multi-target and multi-pathway mechanism of action of HQT in the treatment of UC,providing scientific basis for the clinical development and utilization of HQT.3.HQT can significantly improve the symptoms of intestinal injury and inflammatory infiltration of intestinal mucosa in DSS mice,and its mechanism of action may be through regulating the NLRP3/Caspase-1 signaling pathway,decreasing the expression of NLRP3 and Caspase-1 in colonic tissues,decreasing the expression of pro-inflammatory cytokines IL-6,IL1β and TNF-α in serum,and increasing the expression of anti-inflammatory cytokine IL-10,thus exerting its therapeutic effects.