Mechanism Research Of Qi Bao Mei Ran Dan In Delaying Skin Photoaging Based On Network Pharmacology,Molecular Docking And Experimental Verification

ObejectTo predict the mechanismof Qi Bao Mei Ran Dan in delaying skin photoaging based onnetwork pharmacology and molecular docking,use cell experiments and animal experiments to verify the above-mentioned mechanism of action and signal pathways,confirming Qi Bao Mei Ran Dan resistance from micro to macro Effectiveness of skin photoaging.Methods1.Use TCMSP and TCMID databases to screen the active ingredients and targets of Qi Bao Mei Ran Dan,combine with GeneCards and NCBI Gene databases to obtain skin photoaging-related target genes,build a drug-compound-target network based on Cytoscape software,and STRING database for PPI core Protein interaction analysis,the MCODE plug-in of Cytoscape software performs cluster analysis on the biological functions of core proteins,uses DAVID database to perform GO enrichment analysis and KEGG pathway analysis on 64 core target genes,and uses Autodock Vina,Python and Pymol software to perform Molecular docking,2.In order to verify the effectiveness of Qi Bao Mei Ran Dan against skin photoaging and the mechanism of oxidative stress and collagen proliferation,aging human dermal fibroblasts(NHSF)treated with Qi Bao Mei Ran Dan medicated serum were used to observe cell activity and Refractive performance,to judge the protective effect of Qi Bao Mei Ran Dan on photoaging of human dermal fibroblasts,Western-bolt method to detect inflammatory oxidative stress-related tumor necrosis factor(TNF-α)and collagen-related in human dermal fibroblasts(NHSF)The protein expression level of matrix metalloproteinase(MMP-1),clarify the oxidative stress and collagen proliferation pathways,and the role of MAPK/NF-kB and PI3K/AKT signals in the TNF signaling pathway,verifying the prediction results of network pharmacology.Results1.Obtained 70 effective active ingredients of Qi Bao Mei Ran Dan,273 target points,234 skin photoaging-related target genes,64 cross-core targets,and 393 biological process items obtained by GO enrichment analysis.In oxidative stress,Genes are relatively enriched in biological processes such as apoptosis,lipid metabolism,and angiogenesis.KEGG analysis is represented by TNF signaling pathway.Among them,MAPK/NF-kB signaling pathway and PI3K/AKT signaling pathway have prominent roles,and the core target is TNF,MAPK1,PTGS2,and AKT1 are enriched here to control the gene expression process of skin aging.The molecular docking results showed that the active components of Qi Bao Mei Ran Dan,quercetin,kaempferol,isorhamnetin,daidzein,and the core target proteins AKT1,TNF,PTGS2,MAPK1 molecular docking combined best,and the conformation is stable.2.Qi Bao Mei Ran Dan medicated serum treats aging human dermal fibroblasts(NHSF).The results show that the cells in the model group are arranged irregularly,the cells become flat and enlarged,vacuoles appear,the outline is not clear,the refractive index is poor,and it is in line with aging.Cell morphology characteristics:the Qi Bao Mei Ran Dan group has the same cell morphology,with a long spindle shape,clear outline and good refractive index.The positive rate of SA-β-gal staining in the normal group was(2.78%± 0.22%),the blank group was(2.83% ± 0.29%);the model group was(37.58% ± 2.56%),and the Qi Bao Mei Ran Dan group was(24.55% ±1.74%).Compared with the normal group and the blank group,the positive rate of SA-β-gal staining in the model group was significantly higher,and the difference was statistically significant(P<0.05);compared with the model group,the positive rate of SA-β-gal staining in Qi Bao Mei Ran Dan group was significant Decrease,the difference is statistically significant(P<0.05).Further observation of the expression levels of TNF-αand MMP1 protein in human dermal fibroblasts(NHSF)showed that the model group was compared with the normal group and the blank group,respectively,the expression levels of TNF-α and MMP-1 were significantly increased.There was statistical significance(P<0.05);the expression levels of TNF-α and MMP-1 in the Qi Bao Mei Ran Dan group were significantly lower than the model group,and the difference was statistically significant(P<0.05).TNF-α and MMP1 in each group of NHSF Compared with the normal group and the blank group,the expression levels of TNF-α and MMP-1 in the model group ofprotein were significantly increased;the expression levels of TNF-αand MMP-1 in the Qi Bao Mei Ran Dan group were significantly lower than that of the model group.Conclusion1.Qi Bao Mei Ran Dan may combine with core targets(AKT1,TNF,PTGS2,MAPK1)through core components(quercetin,kaempferol,isorhamnetin,etc.)to regulatethe expression of MAPK/NF-kB and PI3K/AKT signalsin the TNF signaling pathway,then inhibits cell apoptosis,promotes angiogenesis,affects oxidative stress,and collagen metabolism,delays skin photoaging.2.Experiments show that Qi Bao Mei Ran Dan has a significant protective effect on the photoaging of human dermal fibroblasts(NHSF),can regulate cell oxidative stress damage and collagen metabolism,and down-regulate skin tumor necrosis factor TNF-α,matrix metal The expression of protease MMP1 affects the conduction of MAPK/NF-kB and PI3K/AKT signals in the TNF signaling pathway,thereby delaying skin photoaging.This study verified the results of the previous network pharmacology analysis and confirmed that Qi Bao Mei Ran Dan delays the skin part of the mechanism of photoaging.