目的:采用网络药理学-分子对接-实验验证的方法探讨银杏叶提取物金纳多的物质基础与在病理性瘢痕治疗中的潜在机制。方法:在DGIdb、DrugBank、Swiss Target Prediction和TCMSP 数据库中输入关键词“银杏叶”,获得金纳多的作用靶点;从GeneCards、DisGeNET、OMIM和DrugBank数据库获得病理性瘢痕病变疾病相关靶点,取上述靶点交集即为金纳多抗病理性瘢痕作用靶点。将该靶点上传至STRING数据库,所得数据导入Cytoscape 3.9.1,构建蛋白互作网络,筛选出核心作用靶点;使用DAVID数据库对金纳多抗病理性瘢痕作用靶点基因进行GO和KEGG富集分析,对核心作用靶点与金纳多结合力进行分子对接验证,建立小鼠病理性瘢痕模型并验证金纳多抗病理性瘢痕生成的作用。结果:共获得金纳多抗病理性瘢痕作用靶点38个,并筛选出核心靶基因7个,分别为MAPK1、JUN、IL6、FOS、TNF、MAPK14和AKT1。KEGG途径分析富集的信号通路包括:糖尿病并发症中的AGE-RAGE信号通路、脂质和动脉粥样硬化、IL-17信号通路等。分子对接显示,核心靶基因MAPK1、MAPK14和AKT1与金纳多亲和力较好,尤其与槲皮素、木犀草素、山奈酚具有强烈的结合活性。小鼠病理性瘢痕模型给药实验显示,与模型组相比,静脉推注20 mg/(kg·d)金纳多的小鼠病理性瘢痕组织转移生长因子(transforming growth factor-β1, TGF-β1)表达显著下降(P＜0.05)。结论:金纳多抗病理性瘢痕生成具有多靶点、多途径的特点。金纳多主要通过作用于MAPK1、MAPK14及AKT1等核心靶点基因,影响PI3K/AKT/mTOR通路和MAPK信号通路等发挥对病理性瘢痕的治疗作用。
Objective: To explore the material basis of Ginkgo biloba extract Ginaton and its potential mechanism in the treatment of pathological scar by network pharmacology-molecular docking-experimental verification method. Methods: The key word 'Ginkgo biloba' was input into DGIdb, DrugBank, Swiss Target Prediction and TCMSP databases to obtain the target of Ginaton. The disease-related targets of pathological scar lesions were obtained from GeneCards, DisGeNET, OMIM and DrugBank databases, and the intersection of the above targets was taken as the target of Ginaton′s anti-pathological scar effect. The target was uploaded to the STRING database, and the obtained data were imported into Cytoscape 3.9.1 to construct a protein interaction network and screen out the core targets. GO and KEGG enrichment analysis was performed on the target genes of Ginaton′s disease-resistant rational scar using the DAVID database. The binding force between the core target and Ginaton was verified by molecular docking. In addition, a mouse pathological scar model was established to verify the role of Ginaton in resisting pathological scar formation. Results: A total of 38 targets of Ginaton against pathological scar were obtained, and 7 core target genes were screened, which were MAPK1, JUN, IL6, FOS, TNF, MAPK14 and AKT1, respectively. KEGG pathway analysis enriched signaling pathways including: AGE-RAGE signaling pathway in diabetic complications, lipid and atherosclerosis, IL-17 signaling pathway, etc. Molecular docking showed that the core target genes MAPK1, MAPK14 and AKT1 had good affinity with Ginaton, especially with quercetin, luteolin and kaempferol. The administration experiment of pathological scar model in mice showed that compared with the model group, the expression of TGF-β1 in pathological scar tissue of mice injected with 20 mg/(kg·d) Ginaton was significantly decreased (P＜0.05). Conclusion: Ginaton has a multi-target and multi-pathway character in anti-pathological scar generation. Ginaton primarily acts on core target genes such as MAPK1, MAPK14 and AKT1, and affects the PI3K/AKT/mTOR pathway and the MAPK signaling pathway to exert therapeutic effects on pathological scarring.
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