目标 本研究旨在运用网络药理学和分子对接技术,探索苋草合剂外用治疗日光性皮炎的活性成分及其作用机制,并通过动物实验验证其疗效及相关信号通路.方法 实验分为三部分,分别为:(1)活性成分及潜在靶点筛选.通过中药系统药理学数据库(traditional chinese medicine systems pharmacology database and analysis platform,TCMSP)筛选出苋草合剂(由马齿苋和败酱草组成)的活性成分及潜在靶点,再借助GeneCards等数据库筛查与日光性皮炎相关的靶点.(2)关键成分与靶蛋白的结合效果验证.确定药物与疾病的共有靶点,构建并分析蛋白互作网络(protein-protein interaction networks,PPI),并进行基因本体论(gene ontology,GO)和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路的富集分析.并通过分子对接技术验证苋草合剂关键成分与靶蛋白的结合效果.(3)动物在体实验验证.建立紫外线B(ultraviolet B,UVB)辐射诱导的小鼠日光性皮炎模型,UVB辐照强度为0.45 mW/cm2,辐照剂量为810 mJ/cm2,照射时间30 min,照射1次.建模后采用苋草合剂治疗,治疗3 d后,观察小鼠皮肤组织病理学改变,并通过蛋白质印迹法(Western blot,WB)检测关键蛋白的表达水平.结果(1)活性成分及潜在靶点筛选结果:苋草合剂具有14个活性成分及211个潜在靶点,同时鉴定出331个与日光性皮炎相关的靶点,其中49个为药物与疾病共有靶点.(2)关键成分与靶蛋白的结合效果验证结果:PPI网络分析揭示出12个核心靶点.KEGG通路分析结果表明,苋草合剂的作用机制可能通过调控白细胞介素-17(interleukin-17,IL-17)等炎症相关信号通路实现.分子对接结果进一步表明,苋草合剂的活性成分对1L-17通路中的关键靶蛋白[如基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)、趋化因子 CCL2、趋化因子 CXCL8、白细胞介素 6(interleukin-6,IL-6)、白细胞介素 1β(interleukin-1β,IL-1β)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)]具有较强的结合力.(3)动物在体实验验证结果苋草合剂能有效缓解小鼠皮肤损伤,并通过抑制IL-17信号通路中的关键蛋白,显著降低炎症蛋白的表达水平.结论 苋草合剂通过多成分、多靶点的协同作用,能有效缓解小鼠皮肤损伤,并通过抑制IL-17信号通路中的关键蛋白,显著降低炎症蛋白的表达水平.
Efficacy and Mechanism of External Application of Xiancao Mixture in Treatment of Solar Dermatitis Based on Network Pharmacology
Objective To explore the active components and mechanisms of Xiancao mixture in the external treatment of solar dermatitis with network pharmacology and molecular docking techniques and verify its efficacy and related signaling pathways through animal experiments.Methods The experiment was divided into three parts.Firstly,active ingredients and potential targets of Xiancao mixture(comprised of Portulaca oleracea and Patrinia scabiosaefolia)were screened out with the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).Target sites related to solar dermatitis were further identified with such databases as GeneCards.Secondly,the binding effect between key ingredients and target proteins were verified.Shared targets between the drug and disease were determined,and a Protein-Protein Interaction(PPI)network was constructed and analyzed.Enrichment analysis was performed using gene ontology(GO)and the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways.The binding effect of key ingredients in Xiancao mixture with target proteins was confirmed with molecular docking technology.Lastly,in vivo animal experiments were conducted for validation.A mouse model of UVB radiation-induced solar dermatitis was established to evaluate the clinical efficacy of Xiancao mixture,and the expression levels of key proteins were assessed with Western blot.Results(1)Totally 14 active ingredients and 211 potential targets were found in Xiancao mixture via the screening of active ingredients and potential targets.Additionally,331 targets associated with solar dermatitis were identified,of which 49 were found to be common targets shared between the drug and the disease.(2)As to the results of the verification of the binding effects between key ingredients and target proteins,12 core targets was revealed through PPI network analysis.KEGG pathway analysis indicated that the mechanism of action of Xiancao Mixture may operate through the regulation of inflammation-related pathways such as interleukin-17(IL-17)pathway.Molecular docking results further demonstrated strong binding affinities of Xiancao Mixture active ingredients to key target proteins within the IL-17 pathway,including matrix metalloproteinase-9(MMP-9),chemokines CCL2 and CXCL8,interleukin-6(IL-6),interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α).(3)The results of in vivo validation showed that Xiancao Mixture effectively alleviated skin damage in mice and significantly reduced the expression of inflammatory proteins by inhibiting key proteins in IL-17 signaling pathway.Conclusions Xiancao mixture demonstrates the ability to effectively alleviate skin damage in mice through a synergistic effect of multiple components and targets.Additionally,it significantly reduces the expression of inflammatory proteins by inhibiting key proteins in IL-17 signaling pathway.
Solar dermatitisXiancao MixtureNetwork pharmacologyMolecular docking
NETL
NSTL
万方数据
