摘要
目的通过体外实验探究槲皮素和山柰酚对痤疮相关致病菌的抗菌作用,运用网络药理学和分子对接技术预测二者治疗痤疮的作用机制。方法2022年1月至2022年6月通过体外实验测定抑菌环直径、最小抑菌浓度(MIC)和最小杀菌浓度(MBC)、药物时间抑菌曲线评判槲皮素和山柰酚的抗菌活性;检索DrugBank、HERB及BATMAN-TCM数据库收集槲皮素和山柰酚作用靶点,检索GeneCards、OMIM、DisGeNET、DrugBank数据库获得痤疮靶点;取两靶点集交集,通过STRING数据库获得蛋白互作网络,CytoHubba拓扑分析得到核心靶点,Metascape数据库进行基因本体(GO)和京都基因与基因组百科全书(KEGG)通路富集分析,AutoDock软件进行活性成分与核心靶点的分子对接验证。结果槲皮素和山柰酚对三种痤疮相关致病菌均有较好抗菌活性;槲皮素对三种菌的MIC≤78.13 mg/L,MBC≤156.25 mg/L;山柰酚对金黄色葡萄球菌和表皮葡萄球菌的MIC≤39.06 mg/L,MBC≤78.13 mg/L,对痤疮丙酸杆菌的MIC≤78.13 mg/L,MBC≤156.25 mg/L。通过网络药理学筛选出AP-1转录因子亚基(JUN)、转录因子p65(RELA)、肿瘤坏死因子(TNF)、白细胞介素(IL)-6等8个核心靶点;KEGG富集分析示IL-17信号通路、TNF信号通路等7条信号通路发挥关键作用;将两种化合物与JUN、RELA、TNF、IL-6进行对接,结合自由能均小于-20.92 kJ/mol,表明化合物与核心靶点蛋白形成稳定构象。结论槲皮素和山柰酚对三种痤疮致病菌均有抗菌活性;网络药理学研究显示两种中药成分治疗痤疮具有多靶点、多通路协同特点,具有进一步开发和研究的价值。
Objective The antibacterial effects of quercetin and kaempferol on acne-related bacteria were studied in vitro,and the network pharmacology and molecular docking techniques were used to explore the mechanism of the two compounds in the treatment of acne.Methods The antibacterial activity of quercetin and kaempferol was evaluated by measuring the diameter of bacteriostatic ring,minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)and drug time inhibition curve.DrugBank,HERB and BATMAN-TCM databases were searched to collect quercetin and kaempferol targets;GeneCards,OMIM,DisGeNET and DrugBank databases were searched to obtain acne targets;The intersection of the two target sets was selected,and the protein interaction network was obtained by STRING database,the core target was obtained by topological analysis of CytoHubba,and the gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis was conducted by Metascape database.AutoDock software performs molecular docking verification of active ingredients and core targets.Results Quercetin and kaempferol had good antibacterial activity against Staphylococcus aureus,Staphylococcus epidermidis and Cutibacterium acnes.The MIC and MBC of quercetin against acne-related pathogens was≤78.13 mg/L and≤156.25 mg/L,respectively;the MIC and MBC of kaempferol against Staphylococcus aureus and Staphylococcus epidermidis was≤39.06 mg/L and≤78.13 mg/L,respectively.The MIC of Cutibacterium acnes≤78.13 mg/L,MBC≤156.25 mg/L;Bacterial time-growth curve observed that the two drugs had inhibitory effects on the growth of three bacteria.Eight core targets such as AP-1 transcription factor subunit(JUN),transcription factor p65(RELA),tumor necrosis factor(TNF)and interleukin(IL)-6 were screened by network pharmacology;KEGG enrichment analysis showed that seven signal pathways such as IL-17 signal pathway and TNF signal pathway played a key role;docking the two compounds with JUN,RELA,TNF and IL-6,the binding free energies were all less than−20.92 kJ/mol,indicating that the compound formed a stable conformation with the core target protein.Conclusion Quercetin and kaempferol have antibacterial activity against three acne-related pathogens.Network pharmacological studies show that both of them have the characteristics of multi-target and multi-pathway cooperation in the treatment of acne.It has the value of further development and research.
作者
万春梅
石春蕊
高军
张剑虹
WAN Chunmei;SHI Chunrui;GAO Jun;ZHANG Jianhong(The First School of Clinical Medicine,Lanzhou University,Lanzhou,Gansu 730000,China;Department of Dermatology,The First Hospital of Lanzhou University,Lanzhou,Gansu 730000,China)
出处
《安徽医药》
CAS
2023年第10期2092-2098,I0006,共8页
Anhui Medical and Pharmaceutical Journal
基金
甘肃省自然科学基金项目(22JR11RA043)。
