基于网络药理学和分子对接技术探讨淫羊藿–女贞子治疗绝经后骨质疏松症的作用机制

Mechanism of Epimedia-Ligustrum in Treat-ing Postmenopausal Osteoporosis Based on Network Pharmacology and Molecular Docking Technology

摘要: 目的：运用网络药理学和分子对接技术分析淫羊藿–女贞子治疗绝经后骨质疏松的作用机制。方法：利用中药系统药理学数据库与分析平台(TCMSP)筛选淫羊藿–女贞子的有效活性成分及对应靶点。人类基因数据库(Genecards)、在线人类孟德尔遗传数据库(OMIM)、DisGeNET数据库获取绝经后骨质疏松的相关靶点。使用Venn2.1.0在线平台获取药物与疾病的交集靶点，利用STRING在线数据库构建交集靶点的蛋白相互作用关系，并使用Cytoscape3.9.1软件构建网络图。通过Metascape数据库对交集靶点进行GO和KEGG通路富集分析并使用微生信平台绘图，使用Cytoscape3.9.1软件构建淫羊藿–女贞子治疗绝经后骨质疏松的“药物成分–靶点–通路”网络图。最后运用PyMOL等软件对淫羊藿–女贞子的主要活性成分与关键靶点进行分子对接验证。结果：共得到淫羊藿–女贞子活性成分27个，活性成分相关靶点233个；GeneCards、OMIM、DisGeNET数据库得到绝经后骨质疏松相关靶点1083个，经Venn2.1.0在线平台得到交集靶点80个。共筛选到27中有效成分，主要有效成分有：槲皮素(quercetin)、木犀草素(luteolin)、山奈酚(kaempferol)、大黄素(emodin)、脱水淫羊藿素(Anhydroicaritin)等，AKT1 (蛋白激酶1)、IL-6 (白细胞介素-6)、TNF (肿瘤坏死因子)、VEGFA (血管内皮生长因子)、IL-1β (白细胞介素-1β)、MMP9 (基质金属蛋白酶-9)、PTGS2 (前列腺素内过氧化物合酶2)、CCL-2 (淋巴细胞趋化因子-2)、CXCL8 (白细胞介素-8)、EGF (表皮生长因子)，为淫羊藿–女贞子治疗绝经后骨质疏松的核心靶点。GO和KEGG分析结果表明淫羊藿–女贞子治疗绝经后骨质疏松主要涉及细胞分化的负调控、细胞对脂质、脂多糖、细菌来源的反应等生物过程；主要通过参与癌症通路、肿瘤坏死因子信号通路、白细胞介素-17信号通路、PI3K-Akt信号通路、MAPK信号通路等作用于绝经后骨质疏松。分子对接结果表明，羌活胜湿汤中槲皮素、山柰酚、木犀草素等5个主要活性成分与AKT1、IL-6、TNF等5个关键靶点有着较好的亲和力。结论：淫羊藿–女贞子可以通过槲皮素(quercetin)、木犀草素(luteolin)、山奈酚(kaempferol)、大黄素(emodin)、脱水淫羊藿素(Anhydroicaritin)等成分与AKT1 (蛋白激酶1)、IL-6 (白细胞介素-6)、TNF (肿瘤坏死因子)、VEGFA (肿血管内皮生长因子)、IL-1β (白细胞介素-1β)、MMP9 (基质金属蛋白酶-9)、PTGS2 (前列腺素内过氧化物合酶2)、CCL-2 (过淋巴细胞趋化因子-2)、CXCL8 (白细胞介素-8)、EGF (表皮生长因子)等靶点产生相互作用，调节AGE-RAGE信号通路、TNF、PI3K-Akt、GFR等通路实现治疗绝经后骨质疏松的作用。

Abstract: Objective: This paper aims to analyze the mechanism of Epimedia-Ligustrum in treating postmen-opausal osteoporosis by network pharmacology and molecular docking. Methods: The active com-ponents and corresponding targets of Epimedia-Ligustrum were screened using the Systematic Pharmacology Database and Analysis platform (TCMSP). The Human Genetic Database (Genecards), online Human Mendelian Genetic Database (OMIM) and DisGeNET database were used to obtain the relevant targets of postmenopausal osteoporosis. The Venn2.1.0 online platform was used to obtain the intersection targets of drugs and diseases, the protein interaction relationship of intersection targets was constructed using STRING online database, and the network diagram was constructed using Cytoscape3.9.1 software. The GO and KEGG pathway enrichment analysis of intersection tar-gets was carried out through Metascape database, and the mapping was carried out using Weishengxin platform. The “drug component-target-pathway” network map of Epimedia-Ligustrum in the treatment of postmenopausal osteoporosis was constructed using Cytoscape3.9.1 software. Finally, PyMOL software was used to verify the molecular docking between the main active compo-nents and key targets of Fructus herba. Results: A total of 27 active constituents and 23 related targets of active constituents were obtained. 1083 targets related to postmenopausal osteoporosis were obtained from GeneCards, OMIM and DisGeNET databases, and 80 intersecting targets were obtained from Venn2.1.0 online platform. A total of 27 active ingredients were screened, and the main active ingredients were: quercetin, luteolin, kaempferol, emodin, Anhydroicaritin, etc. AKT1 (protein kinase 1), IL-6 (interleukin-6), TNF (tumor necrosis factor), VEGFA (vascular endothelial growth factor), IL-1β (interleukin-1β), MMP9 (matrix metalloproteinase-9), PTGS2 (prostaglandin peroxidase synthase 2), CCL-2 (lymphocyte chemokine-2), CXCL8 (Interleukin-8) and EGF (epider-mal growth factor) are the core targets of Epimedia-Ligustrum in the treatment of postmenopausal osteoporosis. The results of GO and KEGG analysis showed that the treatment of postmenopausal osteoporosis mainly involved the negative regulation of cell differentiation, cell response to lipids, lipopolysaccharides and bacteria. It mainly plays a role in postmenopausal osteoporosis by partici-pating in cancer pathway, tumor necrosis factor signaling pathway, interleukin-17 signaling path-way, PI3K-Akt signaling pathway, MAPK signaling pathway, etc. The molecular docking results showed that quercetin, kaempferol and luteolin in Qiangshengsheng Decoction had good affinity with five key targets such as AKT1, IL-6 and TNF. Conclusion: The protein kinase 1 (AKT1) and IL-6 (leucocytic agent) can be combined with quercetin, luteolin, kaempferol, emodin and Anhydroicari-tin Tin-6), TNF (tumor necrosis factor), VEGFA (vascular endothelial growth factor), IL-1β (interleu-kin-1β), MMP9 (matrix metalloproteinase-9), PTGS2 (prostaglandin peroxidase synthase 2), CCL-2 (hyperlymphocyte chemokine-2), CXCL8 (interleukin-8), EGF (epidermal growth factor) and other targets interact, regulating AGE-RAGE signaling pathway, TNF, PI3K-Akt, GFR and other pathways to achieve the treatment of postmenopausal osteoporosis.