Study on the Mechanism of Yinchenhao Decoction in Treating Diabetes Mellitus Based on Network Pharmacology and Experimental Verification
[Objective]This study was aimed to explore the action mechanism of Yinchenhao decoction(YCHD)in the treatment of diabetes mellitus(DM)by network pharmacology,molecular docking and experimental validation.[Method]TCMSP,ETCM,UniProt,SwissTargetPrediction databases and literature reviews were utilized to find the active ingredients and their related targets.OMIM,PharmGkb,TTD and DrugBank databases were utilized to obtain the disease targets of DM.The protein-protein interaction(PPI)network diagrams of YCHD and DM were constructed,and the key targets were analyzed by GO function and KEGG signal pathway enrichment.Molecular docking was performed using AutoDockTool 1.5.7 software to predict the binding energy of active ingredients and core targets.The cells were divided into control group(Control),model group(Model),and YCHD-containing serum low(YCHD-L),medium(YCHD-M)and high(YCHD-H)dose groups.The levels of inducible nitric oxide synthase(iNOS),cyclooxygenase 2(Cox2),Toll-like receptor 4(TLR4),nuclear factor-KB(NF-κB)p65,interleukin-1β(IL1β),IL6 mRNA and iNOS,COX2,Myd88,NF-κB P65 protein were examined by in vitro cell assay to verify the key pathways.[Result]A total of 147 YCHD active ingredients were obtained,486 DM disease targets were identified,and 57 genes related to the action of YCHD on DM were identified,including insulin(INS)and IL1β,tumor necrosis factor(TNF),nuclear factor kappa-light-chain-enhancer of activated B cells 1(NFKB1),signal transducer and transcriptional activator 1(STAT1),albumin(ALB),TLR4,IL1A,IL10,and IL8 were key target genes.According to the counts value,the top ten entries in GO included 1 biological process,7 cellular components,and 2 molecular functions,which were the positive regulation of RNA polymerase Ⅱ promoter transcription,endoplasmic reticulum membrane,plasma membrane,extracellular region,nucleoplasm,cytosol,cytoplasm,integral component of membrane,identical protein binding,and protein binding.KEGG pathway analysis showed that it was closely related with NOD-like receptors and NF-κB,Toll-like receptors and other inflammatory signaling pathways.The molecular docking results showed that the active ingredients of YCHD had good binding ability with the main targets.The experimental results showed that compared with control group,the mRNA levels of TLR4,iNOS,COX2,NF-κB p65,IL1β,IL6 and the protein expression levels of INOS,COX2,MyD88,NF-κB p65 of cells were significantly increased(P＜0.05)in model group.Compared with model group,the mRNA levels of iNOS,COX2,IL1β,L6 and the protein expression levels of iNOS and COX2 of cells were significantly reduced(P＜0.05)in YCHD-L,YCHD-M and YCHD-H groups.The mRNA levels of TLR4 and NF-κB p65 of cells in YCHD-M and YCHD-H groups were significantly decreased(P＜0.05),and the protein levels of Myd88 and NF-κB p65 of cells in YCHD-H group were significantly decreased(P＜0.05).[Conclusion]YCHD might regulate TLR4,NF-κB and other inflammatory signaling pathways through multiple components and multiple targets to treat the DM.The YCHD-H group showed the most significant inhibition of inflammatory signaling pathway related indicators.
万方数据
维普
