基于网络药理学探讨五参顺脉汤对动脉粥样硬化AopE-/-小鼠的作用机制

doi:10.16431/j.cnki.1671-7236.2023.01.038

摘要/Abstract

摘要： 【目的】预测五参顺脉汤治疗动脉粥样硬化的潜在靶点及作用机制，为五参顺脉汤临床应用提供参考依据。【方法】通过中药系统药理学分析平台（TCMSP）数据库筛选五参顺脉汤抗动脉粥样硬化活性成分及作用靶点，通过GeneCards、OMIM数据库获取动脉粥样硬化靶点并统一使用UniProt数据库进行标准化，使用R语言脚本获取两者交集靶点；采用Cytoscape 3.9.1软件建立药物-成分-靶点网络图并进行拓扑学属性分析；将交集靶点上传至STRING数据库获得蛋白-蛋白互作（PPI）关系网络，并在DAVID平台中进行GO功能和KEGG通路富集分析。采用AopE^-/-雄鼠进行试验验证，HE染色观察主动脉病变情况，ELISA检测单核细胞趋化因子-1（MCP-1）、白细胞介素-6（IL6）、IL1B、肿瘤坏死因子-α（TNF-α）水平。【结果】五参顺脉汤中共有活性成分124个，获得靶点122个，动脉粥样硬化靶点4 711个，两者交集靶点98个。拓扑学分析显示，五参顺脉汤治疗动脉粥样硬化的核心成分为槲皮素、β-谷甾醇、豆甾醇、木犀草素，核心靶点为环氧合酶-1（PTGS1）、类固醇受体激活蛋白2抗体（NOCA2）、猴丝氨酸蛋白酶1（PRSS1）等。GO功能富集分析中生物过程得到321个条目（P<0.01），主要涉及对脂多糖的反应、对肿瘤坏死因子的反应、脂肪酸代谢过程等；分子功能得到93个条目（P<0.01），主要涉及膜筏、膜微域、突触膜等；细胞组分得到45个条目（P<0.01），主要涉及DNA结合转录因子结合、RNA聚合酶Ⅱ特异性DNA结合转录因子结合、核受体的活动等。KEGG通路富集分析中共得到129条信号通路（P<0.01），如癌症通路、脂质和动脉粥样硬化通路、流体剪切应力和动脉粥样硬化通路等，大多与调节免疫性炎症和动脉粥样硬化有关。血脂四项检测结果显示，与模型组相比，药物组小鼠血清中胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）水平显著下降（P<0.05），HDL-C升高（P>0.05）；HE染色结果显示，给药组主动脉斑块明显减少，脂质侵润减轻，泡沫细胞数量减少；ELISA检测结果显示，在药物作用下主动脉组织匀浆中炎症因子MCP-1、IL6、IL1B、TNF-α含量均显著降低（P<0.05）。【结论】五参顺脉汤可通过多成分、多靶点、多通路发挥治疗动脉粥样硬化的作用，其活性成分槲皮素、β-谷甾醇、豆甾醇、木犀草素可能通过作用于PTGS1、乙酰胆碱酯酶（ACHE）、腺苷（AR）等靶点产生炎性控制及免疫调节作用。

关键词: 五参顺脉汤; 网络药理学; 动脉粥样硬化; 血脂; 炎性因子

Abstract: 【Objective】 This study was aimed to predict the potential targets and mechanism of WU Shen venation smooth decoction in the treatment of atherosclerosis, so as to provide reference for the clinical application of WU Shen venation smooth decoction.【Method】 The anti-atherosclerosis active components and targets of WU Shen venation smooth decoction were screened through the database of Traditional Chinese Medicine System Pharmacology Analysis Platform (TCMSP), the atherosclerosis targets were obtained by GeneCards and OMIM databases and standardized by UniProt database, and the intersection targets were obtained by R language script.Cytoscape 3.9.1 software was used to establish the drug-component-target network diagram and analyze the topology properties.Upload the intersection target protein-protein interaction (PPI) relationship network was obtained from STRING database, and GO function and KEGG pathway enrichment analysis were performed in DAVID platform.AopE^-/- male mice were used for experimental verification, HE staining was used to observe the aortic lesions.The levels of monocyte chemokine 1 (MCP-1), interleukin-6 (IL6), IL1B and tumor necrosis factor-α (TNF-α) were determined by ELISA.【Result】 There were 124 active ingredients in WU Shen venation smooth decoction, 112 target genes, 4 711 atherosclerotic targets, and 98 overlapping targets.Topology analysis showed that the key components of WU Shen venation smooth decoction in the treatment of atherosclerosis were quercetin, beta sitosterol, stigmasterol and mignonette.The core targets were prostaglandin-endoperoxide synthase 1 (PTGS1), steroid receptor coactivator 2 (NOCA2), monkey serine protease 1 (PRSS1), etc.In the GO enrichment analysis, biological processes obtained 321 items (P<0.01), mainly related to the response to lipopolysaccharide, tumor necrosis factor, fatty acid metabolism, etc.;Molecular function obtained 93 items (P<0.01), mainly related to membrane rafts, membrane microdomains, synaptic membranes, etc.;Cell component obtained 45 items (P<0.01), which mainly involved DNA-binding transcription factor binding, RNA polymeraseⅡ-specific DNA-binding transcription factor binding, and nuclear receptor activity.A total of 129 signaling pathways (P<0.01) were identified by KEGG enrichment analysis, such as cancer pathway, lipid and atherosclerosis pathway, fluid shear stress and atherosclerosis pathway, etc., and most of them were related to the regulation of immune inflammation and atherosclerosis.The results of four items of blood lipid showed that, compared with model group, the levels of cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in serum of mice in the drug group were decreased significantly (P<0.05), while HDL-C were increased (P>0.05).The results of HE staining showed that the aortic plaques, lipid infiltration and the number of foam cells in the treatment group were significantly reduced.The results of ELISA showed that under the action of drugs, the inflammatory factors MCP-1, IL6, IL1B and TNF-α in aortic tissue homogenate were significantly decreased (P<0.05).【Conclusion】 WU Shen venation smooth decoction could treat atherosclerosis through multiple components, multiple targets and multiple pathways, among which the active ingredients quercetin, β-sitosterol, stigmasterol and luteolin might have inflammatory control and immune regulation effects by acting on PTGS1, acetylcholinesterase(ACHE), adenosine (AR) and other targets.

Key words: WU Shen venation smooth decoction; network pharmacology; atherosclerosis; blood lipid; inflammatory cytokines

中图分类号:

S853.74

刘昊源, 李彦杰, 秦合伟, 金小琴, 牛雨晴, 华晓琼, 张淑芹, 牛丽. 基于网络药理学探讨五参顺脉汤对动脉粥样硬化AopE^-/-小鼠的作用机制[J]. 中国畜牧兽医, 2023, 50(1): 377-389.

LIU Haoyuan, LI Yanjie, QIN Hewei, JIN Xiaoqin, NIU Yuqing, HUA Xiaoqiong, ZHANG Shuqin, NIU Li. Study on the Mechanism of WU Shen Venation Smooth Decoction on Atherosclerosis AopE^-/- Mice Based on Network Pharmacology[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 377-389.

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