In Vitro Antibacterial Effect Of Quercetin And Kaempferol On Acne-related Bacteria And Network Pharmacological Study

Objective To systematically evaluate the efficacy and safety of topical plant preparations in the treatment of mild to moderate acne;explore the antibacterial effects of quercetin and kaempferol on acne-related pathogens（Cutibacterium acnes,Staphylococcus aureus,Staphylococcus epidermidis）through in vitro experiments;use network pharmacology and molecular docking technology to explore the mechanism of the above two compounds in the treatment of acne to provide experimental and theoretical basis for their treatment of acne.MethodsPart one:search Pub Med,The Cochrane Library,Embase,CNKI and VIP databases for randomized controlled trials of topical plant preparations in the treatment of mild to moderate acne from January 2011 to December 2021.After literature screening,data extraction and quality evaluation were carried out independently by two researchers in accordance with the inclusion and exclusion criteria,Meta analysis was carried out by using Rev Man5.3 software.The second part:to investigate the antibacterial activity of quercetin and kaempferol against acne-related pathogens by Agar drilling method,and to evaluate the concentration correlation of different concentrations of drugs against acne-related pathogens;the minimum inhibitory concentration and minimum bactericidal concentration of quercetin and kaempferol were determined by multiple dilution method,and the time bacteriostatic curve was drawn to further investigate their action characteristics.The third part:search Drug Bank,HERB and BATMAN-TCM databases to collect quercetin and kaempferol targets,search Gene Cards,OMIM,Dis Ge NET and Drug Bank databases to obtain acne disease targets;take the intersection of the two target sets,import the intersection targets into STRING database to obtain protein interaction network;use the plug-in Cyto Hubba in Cytoscape software for topological analysis to get the core targets,and Metascape database for GO and KEGG enrichment analysis.Auto Dock software was used to verify the molecular docking between the active components and the core targets.ResultsPart one:1 Compared to the placebo group,the improvement of acne inflammatory lesions in the botanical preparation group was better than that in the placebo group[MD=-1.17,95%CI（-2.25,-0.10）,P<0.05],there was no statistically significant difference between the herbal preparation group and the routine western medicine group（P>0.05）;2 Compared to the placebo group,the improvement of acne non-inflammatory lesion count in the botanical preparation group was better than that in the placebo group[MD=-1.21,95%CI（-2.14,-0.28）,P<0.05],there was no statistically significant difference between the botanical preparation group and the routine western medicine group（P>0.05）;3 Compared to the placebo group,the improvement of total acne lesion count in botanical preparation group was significantly better than that in placebo group[MD=-0.88,95%CI（-1.32,-0.45）,P<0.01],there was no statistically significant difference between botanical preparation group and routine western medicine group（P>0.05）;4 There was no statistically significant difference in the incidence of adverse reactions between the botanical preparation group and the placebo group（P>0.05）.Compared with the conventional western medicine group,the incidence of adverse reactions in the botanical preparation group was significantly less than that in the routine medicine group[RR=0.56,95%CI（0.41,0.78）,P<0.01].The second part:1 Quercetin and kaempferol had good antibacterial activity against Staphylococcus aureus,Cutibacterium acnes,and Staphylococcus epidermidis,and there is a significant positive correlation with drug concentration（r>0.98,P<0.001）;2 The MIC of quercetin against three acne-related pathogens≤78.13μg/m L,MBC≤156.25μg/m L;the MIC of kaempferol to Staphylococcus aureus and Staphylococcus epidermidis≤39.06μg/m L,MBC≤78.13μg/m L,The MIC of Cutibacterium acnes≤78.13μg/m L,MBC≤156.25μg/m L;the MIC of positive control clindamycin to Staphylococcus aureus≤0.13μg/m L,the MIC to Staphylococcus epidermidis≤0.06μg/m L,the MIC of Cutibacterium acnes≤0.03μg/m L;the MIC of erythromycin to Staphylococcus aureus and Staphylococcus epidermidis≤0.13μg/m L,the MIC of Cutibacterium acnes≤0.06μg/m L;3 Through the drug time bacteriostatic curve,it was observed that different concentrations of quercetin and kaempferol had different inhibitory effects on the growth of acne-related pathogenic bacteria.The third part:1 97 drug and disease intersection targets were obtained by network pharmacology,and 8 core targets such as JUN,RELA,TNF and IL6 were further screened;2 GO enrichment analysis showed that cells were mainly involved in biological processes such as lipid response,heterogeneous stimulation response and lipopolysaccharide response.KEGG enrichment analysis showed that seven signal pathways,such as IL-17 signal pathway and TNF signal pathway,played a key role in the treatment of acne with quercetin and kaempferol;3 When the two compounds were docked with JUN,RELA,TNF and IL6,the binding free energies were less than-5 kcal·mol^-1,indicating that the conformational structure of the two compounds with the core target protein was stable and the binding activity was high.Conclusion1 Topical herbal preparations have good curative effect and safety in treating patients with mild to moderate acne.But the conclusions are limited by the quantity and quality of the included studies,and more high-quality,multicenter randomized controlled trials are needed for verification.2 Quercetin and kaempferol had certain antibacterial effects on acne-related pathogens（Staphylococcus aureus,Staphylococcus epidermidis and Cutibacterium acnes）in vitro,and this inhibitory effect was significantly positively correlated with the concentration.3 Comprehensive literature and network pharmacological studies have shown that quercetin plays a role in the inflammatory TNF signal pathway of acne.Kaempferol plays a role by inhibiting reactive oxygen species-dependent MAPKs-NF-κB signal pathway,and they have the characteristics of multi-target and multi-pathway cooperation in the treatment of acne.Through molecular docking,the two compounds form a structurally stable conformation with the core target protein and have high binding activity.The therapeutic effects of quercetin and kaempferol on acne were verified,which has the value of further research and development.