GC-MS结合网络药理学及实验验证挖掘猪牙皂挥发油有效组分及其抗脑缺血再灌注的作用机制  

Effective constituents of essential oil from Gleditsiae Fructus Abnormalis and anti-cerebral ischemia/reperfusion injury mechanism: based on GC-MS, network pharmacology, and experimental verification

作　　者：董娜娜 陈晓兰 邓铋莉 谢树才 胡娟 DONG Na-na;CHEN Xiao-lan;DENG Bi-li;XIE Shu-cai;HU Juan(School of Pharmacy,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China)

机构地区：[1]贵州中医药大学药学院,贵州贵阳550025

出　　处：《中国中药杂志》2023年第4期1076-1086,共11页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金地区基金项目(81860705);贵州省国一流学科(中药学)建设项目(GNYL[2017]008号);贵州省教育厅滚动支持省属高校科研平台团队项目(黔教技[2022]022号)。

摘　　要：基于GC-MS技术,通过网络药理学研究猪牙皂挥发油抗脑缺血再灌注的活性成分、潜在靶点及作用机制,对挖掘出的有效组分进行实验验证。首先,利用GC-MS技术分析明确猪牙皂挥发油成分。其次,通过网络药理学预测成分和疾病靶点,构建药物-成分-靶点的关系网络,并对核心靶点进行基因本体(GO)注释和京都基因与基因组百科全书(KEGG)信号通路富集分析,分子对接模拟活性成分与靶点间的结合活性。最后采用SD大鼠进行实验验证,在成功建立脑缺血再灌注模型的基础上,测定各组大鼠神经学行为评分、脑梗死体积、脑组织病理学形态,ELISA法测定白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量,Western blot法测定血管内皮生长因子(VEGF)蛋白的表达。网络药理学筛选得到22个活性成分、17个核心靶点和56个GO条目,KEGG通路主要有TNF信号通路、VEGF信号通路和鞘脂信号通路等,分子对接结果表明活性成分与靶点亲和力良好,动物实验结果显示猪牙皂挥发油有效组分能降低大鼠神经功能损伤,减少脑梗死体积和IL-1β、IL-6、TNF-α的含量,下调VEGF的表达,验证了网络药理学部分预测结果。该研究体现了猪牙皂挥发油多成分、多靶点、多途径的作用特点,其有效组分抗脑缺血再灌注损伤的机制与TNF、VEGF通路有关,为猪牙皂的深度研究与二次开发提供了新的方向。Based on GC-MS and network pharmacology, the active constituents, potential targets, and mechanism of essential oil from Gleditsiae Fructus Abnormalis(EOGFA) against cerebral ischemia/reperfusion(I/R) injury were explored, and the effective constituents were verified by experiment. To be specific, GC-MS was used identify the constituents of the volatile oil. Secondly, the targets of the constituents and disease were predicted by network pharmacology, and the drug-constituent-target network was constructed, followed by Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment of the core targets. Molecular docking was performed to investigate the binding affinity between the active constituents and the targets. Finally, SD rats were used for experimental verification. The I/R injury model was established, and the neurological behavior score, infarct volume, and pathological morphology of brain tissue were measured in each group. The content of interleukin-1β(IL-1β), interleukin-6(IL-6), and tumor necrosis factor-alpha(TNF-α) was determined by enzyme-linked immunosorbent assay(ELISA), and the protein expression of vascular endothelial growth factor(VEGF) by Western blot. A total of 22 active constituents and 17 core targets were screened out. The core targets were involved in 56 GO terms and the major KEGG pathways of TNF signaling pathway, VEGF signaling pathway, and sphingolipid signaling pathway. Molecular docking showed that the active constituents had high affinity to the targets. The results of animal experiment suggested that EOGFA can alleviate the neurological impairment, decrease the cerebral infarct volume and the content of IL-1β, IL-6 and TNF-α, and down-regulate the expression of VEGF. The experiment verified the part results of network pharmacology. This study reflects the multi-component, multi-target, and multi-pathway characteristics of EOGFA. The mechanism of its active constituents is related to TNF and VEGF pathways, which provides a new directi

关 键 词：猪牙皂挥发油 脑缺血再灌注 GC-MS 网络药理学 分子对接 有效组分 机制 

分 类 号：R285.5[医药卫生—中药学]