背景:黄芪甲苷(AST-IV),作为黄芪的有效活性成分(Fisch。)邦奇。已经发现AST-IV抑制登革热病毒的复制,乙型肝炎病毒,腺病毒,和柯萨奇病毒B3.肠道病毒71型(EV71)是重症手足口病(HFMD)的主要病原,但没有特效药.在这项研究中,我们重点研究AST-IV是否能抑制EV71的复制,并探讨其潜在的潜在机制.
方法:用EV71感染GES-1或RD细胞,用AST-IV处理,或与EV71和AST-IV共同处理。EV71结构蛋白VP1水平,使用蛋白质印迹和50%组织培养感染剂量(TCID50)测量上清液中的病毒滴度,分别。网络药理学用于预测AST-IV抑制EV71复制的可能途径和靶标。此外,采用超高效液相色谱-高分辨质谱(UHPLC-HRMS)技术研究了AST-IV潜在的靶向代谢产物。代谢物和表观指标之间的关联通过Spearman算法进行。
结果:本研究表明AST-IV能有效抑制EV71的复制。网络药理学提示AST-IV通过靶向PI3K-AKT抑制EV71复制。代谢组学结果表明,AST-IV通过提高次黄嘌呤的水平来达到这些效果,2-酮丁酸,腺嘌呤,烟酸单核苷酸,前列腺素H2,6-羟基-1H-吲哚-3-乙酰胺,氧普尿酸,同时降低PC的水平(14:0/15:0)。此外,AST-IV还通过降低MDA水平减轻EV71诱导的氧化应激,ROS,在增加T-AOC活性的同时,CAT,GSH-Px.当使用ROS抑制剂N-乙酰半胱氨酸(NAC)时,也观察到EV71复制的抑制。此外,AST-IV显示出激活PI3K-AKT信号通路并抑制EV71诱导的细胞凋亡的能力。
结论:这项研究表明,AST-IV可能通过靶向八种关键代谢产物激活cAMP和抗氧化应激反应,包括次黄嘌呤,2-酮丁酸,腺嘌呤,烟酸单核苷酸,前列腺素H2,6-羟基-1H-吲哚-3-乙酰胺,氧代嘌呤醇和PC(14:0/15:0)。这种激活可以进一步刺激PI3K-AKT信号传导以抑制EV71诱导的细胞凋亡和EV71复制。
BACKGROUND: Astragaloside IV (AST-IV), as an effective active ingredient of Astragalus membranaceus (Fisch.) Bunge. It has been found that AST-IV inhibits the replication of dengue virus, hepatitis B virus, adenovirus, and coxsackievirus B3. Enterovirus 71 (EV71) serves as the main pathogen in severe hand-foot-mouth disease (HFMD), but there are no specific drugs available. In this study, we focus on investigating whether AST-IV can inhibit EV71 replication and explore the potential underlying mechanisms.
METHODS: The GES-1 or RD cells were infected with EV71, treated with AST-IV, or co-treated with both EV71 and AST-IV. The EV71 structural protein VP1 levels, the viral titers in the supernatant were measured using western blot and 50% tissue culture infective dose (TCID50), respectively. Network pharmacology was used to predict possible pathways and targets for AST-IV to inhibit EV71 replication. Additionally, ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) was used to investigate the potential targeted metabolites of AST-IV. Associations between metabolites and apparent indicators were performed via Spearman\'s algorithm.
RESULTS: This study illustrated that AST-IV effectively inhibited EV71 replication. Network pharmacology suggested that AST-IV inhibits EV71 replication by targeting PI3K-AKT. Metabolomics results showed that AST-IV achieved these effects by elevating the levels of hypoxanthine, 2-ketobutyric acid, adenine, nicotinic acid mononucleotide, prostaglandin H2, 6-hydroxy-1 H-indole-3- acetamide, oxypurinol, while reducing the levels of PC (14:0/15:0). Furthermore, AST-IV also mitigated EV71-induced oxidative stress by reducing the levels of MDA, ROS, while increasing the activity of T-AOC, CAT, GSH-Px. The inhibition of EV71 replication was also observed when using the ROS inhibitor N-Acetylcysteine (NAC). Additionally, AST-IV exhibited the ability to activate the PI3K-AKT signaling pathway and suppress EV71-induced apoptosis.
CONCLUSIONS: This study suggests that AST-IV may activate the cAMP and the antioxidant stress response by targeting eight key metabolites, including hypoxanthine, 2-ketobutyric acid, adenine, nicotinic acid mononucleotide, prostaglandin H2, 6-Hydroxy-1 H-indole-3-acetamide, oxypurinol and PC (14:0/15:0). This activation can further stimulate the PI3K-AKT signaling to inhibit EV71-induced apoptosis and EV71 replication.