基于网络药理学及实验验证探讨玉肤解毒膏治疗卡培他滨所致手足综合征的作用机制

Network Pharmacology-Based Exploration of the Mechanism of Action and Experimental Validation of Yufu Jiedu Ointment(玉肤解毒膏)in the Treatment of Capecitabine-Induced Hand-Foot Syndrome

目的:基于网络药理学与实验验证探讨玉肤解毒膏治疗卡培他滨所致手足综合征(HFS)的作用机制。方法:采用高效液相色谱仪鉴定玉肤解毒膏化学成分;通过TCMSP、PubChem、Swiss Target Prediction数据库筛选玉肤解毒膏活性成分及作用靶点;通过String平台进行蛋白互作网络分析;GeneCards数据库筛选HFS相关靶点;使用Metascape数据库对交集靶点进行基因本体(GO)富集功能分析、京都基因与基因百科组全书(KEGG)通路富集分析;运用AutoDocktools软件对活性成分及核心靶点进行分子对接验证。构建HFS大鼠模型,通过蛋白免疫印迹法检测比较玉肤解毒膏和尿素软膏对HFS大鼠足跖部皮肤中组蛋白去乙酰化酶6(HDAC6)、Toll样受体4(TLR4)蛋白表达水平的影响。结果:共筛选得到玉肤解毒膏活性成分97个,成分作用靶点304个,与HFS交集靶点45个。关键靶点主要富集在Toll样受体信号通路、中性粒细胞胞外陷阱形成、Rap1通路等信号通路。活性成分龙血素A、漆黄素、乔松素、紫檀芪、芫花素与核心靶点HDAC6、组蛋白去乙酰化酶2(HDAC2)、组蛋白去乙酰化酶1(HDAC1)、组蛋白去乙酰化酶4(HDAC4)、信号转导和转录激活因子3(STAT3)、组织蛋白酶D(CTSD)具有较好的结合活性和稳定性。玉肤解毒膏可显著降低HFS大鼠足跖部皮肤中HDAC6、TLR4蛋白表达水平。结论:玉肤解毒膏可能通过调控HDAC6、TLR4表达而降低神经炎症反应,发挥治疗HFS的作用。

Objective:To exploring the mechanism of Yufu Jiedu ointment in the treatment of capecitabine-induced hand foot syndrome(HFS)based on network pharmacology and in vivo experiment validation.Methods:UPLC-Q-TOF-MS was used to identify the chemical constituents of Yufu Jiedu ointment.TCMSP,PubChem and Swiss target prediction databases were used to screen the active ingredients and targets of Yufu Jiedu ointment.String platform was used to analyze the protein interactions.GeneCards database was used to screen the targets related to HFS.GO functional analysis and KEGG signaling pathway enrichment analysis were carried out according to Metascape database.And AutoDocktools software was used to validate the molecular docking of the active ingredients and the core target points.A rat model of HFS was constructed,and Western blotting was used to detect the effects of Yufu Jiedu ointment and urea ointment on the protein expression levels of histone deacetylase 6(HDAC6)and toll-like receptor 4(TLR4)in the plantar skin of HFS rats.Results:A total of 97 active ingredients,304 targets and 45 targets intersecting with HFS were obtained from the screening of Yufu Jiedu ointment.The key targets were mainly enriched in Toll-like receptor signaling pathway,Neutrophil extracellular trap formation,Rap1 pathway and other signaling pathways.The active ingredients,Loureirin A,Fisetin,Pinocembrin,Pterostilbene and Genkwanin,have good binding activity and good stability with the core targets,including HDAC6,histone deacetylase 2(HDAC2),histone deacetylase 1(HDAC1),histone deacetylase 4(HDAC4),signal transducer and activator of transcription 3(STAT3)and cathepsin D(CTSD).In vivo experiments showed that Yufu Jiedu ointment significantly reduced the levels of HDAC6 and TLR4 protein expression in the plantar skin of HFS rats.Conclusion:Yufu Jiedu ointment may reduce neuroinflammatory responses and exert therapeutic effects on HFS by modulating HDAC6 and TLR4 expression.