基于网络药理学和动物实验探讨复方红花补蒺颗粒治疗白癜风的作用机制

Role and mechanism of Fufang Honghua Buji granules in the treatment of vitiligo based on network pharmacology and animal experiments

目的运用网络药理学和体内实验探究复方红花补蒺颗粒(FHB)治疗白癜风的作用机制。方法通过中药系统药理学分析平台(TCMSP)数据库和本草组鉴(HERB)数据库获取红花补蒺方的化学成分和作用靶点。通过在线人类孟德尔遗传数据库(OMIM)、DrugBank数据库、DisGeNET数据库,GeneCards数据库和治疗目标数据库(TTD)筛选白癜风(vitiligo)的相关靶点。借助Matascape数据库进行基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析。利用Cytoscape及STRING数据库构建红花补蒺方“活性成分-靶点-疾病”网络、蛋白相互作用网络,根据拓扑学参数筛选红花补蒺方治疗白癜风的核心成分及作用靶点。最后,将小鼠分为空白组、白癜风模型组、红花补蒺方低剂量组(HQ+0.8 g/kg FHB组)、红花补蒺方中剂量组(HQ+2.4 g/kg FHB组)、红花补蒺方高剂量组(HQ+7.2 g/kg FHB组)。利用酶联免疫吸附实验(ELISA)检测各组小鼠血清中肿瘤坏死因子-α(TNF-α)、胆碱酯酶(CHE)、白介素-6(IL-6)和酪氨酸酶(TYR)的表达,苏木精-伊红染色(HE)实验检测病变皮肤中表皮增生程度,毛囊和黑色素颗粒的数量变化。结果网络药理学结果表明复方红花补蒺颗粒中的核心成分为木犀草素(luteolin)、汉黄芩素(wogonin)和槲皮素(quercetin),其核心治疗靶点为RELA、STAT3和AKT1。此外,复方红花补蒺颗粒可能通过JAK-STAT和NF-κB等信号通路共同治疗白癜风。HE染色结果显示,与模型组相比,HQ+0.8 g/kg FHB组、HQ+2.4 g/kg FHB组和HQ+7.2 g/kg FHB组小鼠病变皮肤表皮均增生减轻,毛囊和黑色素颗粒均增加。ELISA结果显示,与模型组相比,HQ+0.8 g/kg FHB组、HQ+2.4 g/kg FHB组和HQ+7.2 g/kg FHB组小鼠血清中TNF-α、IL-6和CHE含量均显著降低(P<0.05),TYR含量均显著升高(P<0.01)。结论复方红花补蒺颗粒通过多成分、多靶点、多通路结合来发挥抗炎、促黑的作用。

Objective To investigate the mechanism of Fufang Honghua Buji granules(FHB)in the treatment of vitiligo using network pharmacology and in vivo experiments.Methods The main active components and targets of FHB were screened by searching the TCMSP database and HERB database.The relevant targets of vitiligo were searched by OMIM database,DrugBank database,DisGeNET database,GeneCards database,and therapeutic target database(TTD).Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were performed based on Matascape database.The“active ingredient-target-disease”network and the protein interaction network of FHB were constructed by Cytoscape and STRING database,and the core components and targets of FHB in the treatment of vitiligo were screened according to topology parameters.Finally,the mice were divided into blank group,vitiligo model group,FHB low dose group(HQ+0.8 g/kg FHB),FHB medium dose group(HQ+2.4 g/kg FHB),and FHB high dose group(HQ+7.2 g/kg FHB).The serum levels of tumor necrosis factor-α(TNF-α),cholinesterase(CHE),interleukin-6(IL-6),and tyrosinase(TYR)were detected by enzyme-linked immunosorbent assay(ELISA).And the changes of epidermal proliferation,hair follicles and melanin granules in the lesioned skin were observed by hematoxylin-eosin staining.Results The network pharmacology results indicated that the core components in FHB were luteolin,wogonin and quercetin,and their core therapeutic targets were RELA,STAT3,and AKT1.In addition,FHB might treat vitiligo through JAK-STAT and NF-κB-class signaling pathways.HE staining results showed that compared with model group,the hyperplasia of lesioned skin epidermis reduced,and hair follicles and melanin granules increased in HQ+0.8 g/kg FHB group,HQ+2.4 g/kg FHB group,and HQ+7.2 g/kg FHB group.The serum levels of TNF-α,IL-6 and CHE in HQ+0.8 g/kg FHB,HQ+2.4 g/kg FHB,and HQ+7.2 g/kg FHB groups were significantly lower than those in model group(P<0.05),while TYR level was significantly higher(P<0.01).Conclusion FHB can play the anti-inflammatory and pro-black effects through the multiple components and the multiple targets and pathways.