The Mechanism of Yupingfeng Granules(玉屏风颗粒)in Regulating Th1/Th2 Balance in Mice with IgA Nephropathy Model Based on Network Pharmacology
Objective:To predict the mechanism of Yupingfeng granules for the treatment of IgA nephropathy(IgAN)based on network pharmacology and to perform mechanism validation.Methods:The active ingredients of Yupingfeng granules were retrieved from TCMSP database.The IgAN targets were obtained from GeneCards,OMIM and TTD databases,and the intersecting targets of Yupingfeng granules and IgAN were obtained by using the software of Venny 2.1.0 and Cytoscape 3.7.2 to construct the"active ingredient-intersecting target".DAVID database was used to carry out GO and KEGG enrichment analysis.String database was used to construct protein-protein interaction network diagram,and AutoDock Vina software was applied to validate the molecular docking between the main active ingredients and the key targets of Yupingfeng granules.The molecular docking verification was carried out by AutoDock Vina software.Totally 60 male Kunming mice were randomly divided into normal group(n=10)and modelling group(n=50).After successful modelling,the mice were randomly divided into Yupingfeng granules high,medium and low groups,and western medicine group.The mice were administered for eight weeks.Immunofluorescence method was used to detect the deposition of IgA in renal tissues;HE staining and glycogen staining were used to observe the pathological changes of renal tissues;Biochemical method was used to detect 24 h-UTP,BUN and Scr;ELISA method was used to detect the expression of IgA,Th1-related factors,including TNF-α,INF-γ and IL-2,and the expression of Th2-related factors,including IL-4 and IL-6.Results:In the cyberpharmacology part,34 active ingredients of Yupingfeng granules were screened,and there were 222 drug targets,1 436 disease targets,and 102 intersecting targets.The main active ingredients were quercetin,kaempferol,wogonin and so on.The key targets were TNF,IL-6,IL-4,IL-2 and so on.The results of molecular docking showed that the main active ingredients and key targets could be well combined,and the binding energies were≤-6.0 kJ/mol;GO enrichment analysis obtained 689 biological processes,54 cellular components and 115 molecular functions;KEGG enrichment analysis obtained IL-17,NF-κB and other signaling pathways.In animal experiments,IgA immunofluorescence of renal tissues showed that compared with normal group,the model group showed strong green IgA fluorescence signals in the mesangial area,and that compared with model group,the IgA deposition improved in each treatment group,among which,the improvement was more obvious in Yupingfeng granules high dose group.HE staining showed that compared with normal group,the model group showed glomerular swelling,proliferation of mesangial cells and endothelial cells,increase in extracellular matrix,and expansion of the mesangial area,and that compared with model group,the pathological damage of renal tissue was reduced in each treatment group.Glycogen staining showed that compared with normal group,model group showed proliferation of mesangial cells and increase in extracellular matrix,and that the degree of renal pathological changes of treatment groups changed from moderately severe to moderately mild after the administration of the drug.Compared with the normal group,24 h-UTP,BUN and Scr increased in model group(P＜0.01).Compared with model group,renal function was improved in treatment groups(P＜0.01 or P＜0.05).Compared with normal group,IgA expression in serum increased in model group.Compared with model group,IgA expression in serum was reduced in treatment groups(P＜0.05).Compared with normal group,the expression levels of Th1-related factors,including TNF-α,INF-γ,and IL-2,were significantly reduced in model group(P＜0.05),and the Th1-related factors,including TNF-α,INF-γ and IL-2 were up-regulated in treatment groups.Compared with normal group,the expression levels of Th2-related factors,including IL-4 and IL-6,increased in model group(P＜0.05),and the expression levels of Th2-related factors,including IL-4 and IL-6 were down-regulated after the use of drug treatment in treatment groups.Compared with normal group,the serum Th1/Th2 ratio was significantly decreased in model group(P＜0.05),and the serum Th1/Th2 ratio was up-regulated in treatment groups(P＜0.05).Conclusion:Yupingfeng granules have better therapeutic effect on IgAN model mice,and the mechanism may be related to the correction of Th1/Th2 imbalance.
IgA nephropathyYupingfeng granulesTh1/Th2network pharmacologyexperimental validationmice
万方数据
维普
