目的 基于网络药理学研究银杏叶提取物防治老年性聋的的潜在作用机制，并通过动物实验进行初步验证。方法 运用中药系统药理学分析平台（TCMSP）筛选银杏叶的有效成分，GeneCards数据库获得老年性聋相关靶点基因，微生信对基因本体论（GO）、基因组百科全书（KEGG）进行功能富集，Cytoscape3.8.2构建疾病-靶点-通路网络图。采用D-半乳糖诱导SD大鼠制备老年性聋的动物模型，腹腔注射低、中、高剂量（10、20、30 mg·kg^-1）的银杏叶提取物，运用免疫荧光染色耳蜗基底膜铺片观察毛细胞形态变化；透射电子显微镜观察螺旋神经节细胞及线粒体超微结构变化；酶联免疫吸附实验（ELISA）检测血清谷胱甘肽S转移酶（GST）、总超氧化物歧化酶（T-SOD）、丙二醛（MDA）含量；免疫印迹法检测Bcl-2、Bax蛋白表达及Bcl-2/Bax比值变化。结果 GO富集主要涉及细胞衰老、过氧化物酶成分、氧化还原酶的活性等；KEGG富集相关性较高的通路包括细胞凋亡通路和氧化应激相关的谷胱甘肽代谢通路，与凋亡相关的靶点基因包括：Bcl-2、Bax、AKT1、TNF等；与谷胱甘肽代谢相关靶点基因涉及到：GSTP1、GSTM1等。动物实验证实银杏叶提取物可减轻毛细胞丢失，降低螺旋神经节细胞及线粒体损伤，提高GST、T-SOD含量，降低MDA含量，上调Bcl-2表达和Bcl-2/Bax比值，下调Bax表达。结论 银杏叶提取物可能通过Bcl-2/Bax通路减轻耳蜗螺旋神经节细胞和毛细胞的氧化应激和细胞凋亡来延缓老年性聋的进程。

Objective To study the potential mechanism of Ginkgo biloba extract in preventing and treating presbycusis based on network pharmacology, and to conduct preliminary verification through animal experiments.Methods Traditional Chinese Medicine Systems Pharmacology Analysis Platform (TCMSP) was used to screen the active ingredients of Ginkgo biloba. The GeneCards database was used to obtain the target genes related to presbycusis; missense performed functional enrichment on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genome (KEGG); and Cytoscape 3.8.2 constructed a disease-target-pathway network diagram. D-galactose was used to induce SD rats to prepare animal models of presbycusis. Low, medium and high doses (10, 20, 30 mg·kg^-1) of Ginkgo biloba extract were injected intraperitoneally. The morphological changes of hair cells were observed by immunofluorescence staining of cochlear basement membrane paving. The ultrastructural changes of spiral ganglion cells and mitochondria were observed by transmission electron microscopy. The contents of serum glutathione S-transferase (GST), total superoxide dismutase (T-SOD) and malondialdehyde (MDA) were detected by enzyme-linked immunosorbent assay (Elisa). The expression of Bcl-2 and Bax protein and the change of Bcl-2/Bax ratio were detected by Western blotting.Results GO functions were mainly enriched in cell senescence, peroxidase composition and oxidoreductase activity, etc. The KEGG signaling pathway mainly focused on apoptosis pathway stress-related glutathione metabolism pathway, etc. And apoptosis-related target genes included Bcl-2, Bax, AKT1, TNF, etc. Target genes related to glutathione metabolism include GSTP1 and GSTM1 etc. Animal experiments confirmed that Ginkgo biloba extract can reduce hair cell loss, reduce spiral ganglion cells and mitochondrial damage, increase the content of GST and T-SOD, reduce the content of MDA, up-regulate the protein expression of Bcl-2 and Bcl-2/Bax ratio, down-regulate the protein expression of Bax expression.Conclusion The Ginkgo biloba extract may reduce oxidative stress and apoptosis of cochlear spiral ganglion cells and hair cells through Bcl-2/Bax pathway to delay the process of Presbycusis.

国家自然科学基金委员会青年项目（81403439）：基于中医脾肾相关理论探讨ENaC和AQP在梅尼埃病水钠转运中的作用机制，负责人：郭向东；河南省科学技术厅科技攻关计划（192102310170）：补肾活血汤及拆方通过Sestrin2-AMPK-p70S6K信号通路对老年性聋的防护作用，负责人：郭向东；河南省中医药管理局河南省中医药科学研究专项重点课题（20-21ZY1045）：补肾活血汤调控miRNA-29a防治老年性聋的机制与临床研究，负责人：郭向东。