Network Pharmacology Approach To Explore The Skin-lightening Compounds And Potential Mechanisms Of Chinese Herbal Medicines

Purpose: 1.Search for lightening active ingredients in traditional Chinese herbal medicines using network pharmacology techniques.2.Explorelightening mechanisms of potential active ingredients.Methods: 1.Network pharmacology analysis: Use network pharmacology technology to screen potential traditional Chinese herbs and active ingredients that inhibit skin pigmentation in ancient formulas,and perform PPI analysis and KEGG enrichment of potential target genes of active ingredients to explore the whitening mechanism of the active ingredients.2.Culture of melanin-rich human melanoma cells(MNT1)and immortalized human keratinocytes(HaCaT)with and without drug intervention by candidate active ingredients: CCK8 method to detect cell proliferation activity;Fontana Masson method to observe MNT1 cells.Melanin situation;qRT-PCR detected the expression levels of melanin synthesis-related genes in MNT1 cells and paracrine inflammatory factors(ET-1,VEGF,PTGS2,etc.)in HaCaT cells.3.Use molecular docking technology to explore molecules that may directly act on candidate active ingredients.4.After the candidate active ingredient drug intervenes in the 3D model of the skin,the Fontana Masson method is used to detect the melanin in the model.Results: 1.Eight potential whitening Chinese herbs(Ampelopsis Japonica,Schizonepetae Herba,Scutellariae Radix,Asari Radix Et Rhizoma,Semen Pharbitidis,A.Dahurica(Fisch.)Benth.Et Hook,Angelicae Sinensis Radix,Persicae Semen)were screened from ancient formulas through network pharmacology technology,and 10 potential whitening active ingredients such as Quercetin and Acacetin were identified.2.PPI analysis found that the TOP10 core target genes of quercetin are JUN,AKT1,IL6,CASP3,IL1 B,VEGFA,MMP9,TNF,PTGS2,EGF;the core target genes of acacia are TP53,CDKN1 A,HSP90AA1,PTGS2,CASP3,RELA,CASP8.The KEGG enrichment results suggest that Quercetin may regulate signaling pathways such as NF-κB,while Acacetin may regulate signaling pathways such as P53.3.In the low concentration range,Quercetin(0,5,10,20 μmol/L)and Acacetin(0,5,10 μmol/L)alone had no significant effect on melanin synthesis in MNT1 cells;4.After Quercetin and Acacetin were treated in the co-culture system of MNT1 and HaCaT,the level of melanin and the mRNA levels of melanin synthesis-related genes decreased;5.The supernatant of HaCaT cells treated with Quercetin and Acacetin can reduce the content of melanin in MNT1 cells;6.Quercetin and Acacetin can inhibit the mRNA levels of paracrine inflammatory factors(ET-1,VEGF,PTGS2,etc.)in HaCaT cells.7.Molecular docking indicated that Quercetin and Acacetin had strong affinity for the key proteins TLR4,CHUK and RELA in the NF-κB pathway.8.Quercetin and Acacetin can inhibit melanin synthesis in skin 3D models.Conclusion: 1.The network pharmacology screening indicated that Chinese herbal medicines such as Ampelopsis Japonica,Schizonepetae Herba,Scutellariae Radix,Asari Radix Et Rhizoma,Semen Pharbitidis,A.Dahurica(Fisch.)Benth.Et Hook,Angelicae Sinensis Radix,Persicae Semen have potential whitening effects,while Quercetin and Acacetin are potential whitening active ingredients.2.Quercetin and Acacetin can inhibit melanogenesis by affecting the paracrine effect of keratinocytes.3.Quercetin and acacia may inhibit the paracrine function of keratinocytes by inhibiting multiple targets of NF-κB signaling pathway.