Objective:This study aims to reveal the common regulatory network of Transcription Factor(TF)in AD and rosacea,predict the potential drugs and pharmacological targets that can treat AD and rosacea simultaneously,and verify them experimentally.Methods:1)The prediction of shared regulatory network and potential drug for both AD and rosacea:The differentially expressed genes(DEGs)in AD and rosacea were obtained by differential and intersection analysis of transcriptome sequencing data of AD and rosacea and related control tissues.The common TFs regulatory network of both AD and rosacea was constructed by analyzing these DEGs with bioinformatics technology.The potential drugs for treating both AD and rosacea were predicted by network pharmacology.Melatonin(MLT)has been proven to treat AD,so we only verified the therapeutic effect of MLT in rosacea.2)Experimental verification of the efficacy of MLT on rosacea:MLT was used to treat rosacea-like mouse model induced by LL37,and the effects of MLT on the phenotype of rosacea in mice were evaluated by measuring the skin thickness,skin area and erythema degree of each group.The effect of MLT on the number of immune cells and neovascularization in rosacea lesions was detected by immunofluorescence.Using q PCR to assess the effect of MLT on m RNA expression of rosacea related factors in rosacea-like mouse skin tissues and Human immortalized keratinocyte(Ha Ca T)induced by LL37/TNF-α.The inhibitory effect of MLT on nuclear factor kappa-B(NF-κB)activation was evaluated by cell fluorescence and Western-blot.Through the cell migration experiment and cell chemotaxis experiment to assess the effect of MLT on chemotaxis and migration ability of human umbilical vein endothelial cells(HUVEC)induced by LL37.3)The pharmacological targets of MLT for treating both AD and rosacea:The public pharmacological database was used to predict the pharmacological targets of MLT,and the intersection analysis of the common TFs-target genes with AD and Rosacea was conducted to identify the pharmacological targets of MLT for the treatment of AD and Rosacea.The functional and pathway enrichment analysis was conducted to determine the biological processes and courses related to these target genes.The Protein-Protein Interaction(PPI)network of these drug targets was constructed,and the key targets with the highest correlation were selected.The binding of MLT to these key targets was analyzed by molecular docking.Results:1)TFs regulatory network analysis and prediction of potential therapeutic drugs for AD and rosacea:Through DEGs analysis of transcriptome sequencing results,DEGs of 5090 AD patients and 3016 rosacea patients were obtained.The 747 shared DEGs of AD and rosacea were determined by intersection analysis.Through enrichment and cluster analysis,it was found that these biological processes,pathways and diseases with shared DEGs enrichment were related to inflammation and vascular regulation.These shared DEGs and Trust databases were used to construct AD and rosacea TFs regulatory networks.MLT was predicted to be a potential drug for the treatment of AD and rosacea through network pharmacology.2)The mechanism of MLT in the treatment of rosacea:MLT significantly improved LL37-induced rosacea-like phenotype,decreased the number of CD4~+T cells,F4/80~+cells and CD31~+vascular density in rosacea-like mouse skin lesions,and inhibited the m RNA expression of rosacea-related factors in Ha Ca T cells induced by LL37/TNF-α.MLT can effectively block the activation of NF-κB in Ha Ca T cells induced by TNF-αand inhibit angiogenesis and migration and chemotaxis of HUVECs induced by LL37.3)Pharmacological target study of MLT treating of AD and rosacea:19 pharmacological targets of MLT in treating AD and rosacea were revealed through network pharmacological analysis.Through molecular docking of MLT with 8 key targets with the highest correlation,it was found that MLT had a good binding effect with CCND1,EGFR,ICAM1,MMP-9,SERPINE1 and PTGS2.Conclusions:For the first time,this study reveals that AD and rosacea shared a TFs regulatory network and predicts and verifies that MLT is a potential drug for treating AD and rosacea.Eight pharmacological targets of MLT in the treatment of AD and rosacea were identified by molecular docking. |