背景:登革热,由登革热病毒引起的(正氟病毒登革热,包括DENV类型1-4),是黄病毒科的一员.登革热的症状从亚临床或轻度表现到潜在的致命并发症。由于缺乏有效的抗病毒药物,严重登革热的管理非常具有挑战性。在这种情况下,天然产品,无论是纯化合物还是标准化植物提取物的形式,已成为开发新的抗病毒疗法的有希望的来源。Hernandonine,在HernandiaNymphaifolia中发现的一种氧杂吗啡生物碱(C.Presl)库比茨基。作为人类免疫缺陷病毒1型(HIV-1)整合酶的代谢物和抑制剂。
目的:本研究探讨了西恩多宁抑制DENV感染的能力,并探讨了其潜在机制。
方法:为了评估体外抗DENV活性,在用DENV感染之前或之后,将细胞或诱导多能干细胞(iPSC)来源的脑类器官暴露于Hernandonine。和Hernandonine一起,胞吞调节剂,Genistein,Wortmannin,甲基-β-环糊精(MβCD)和洛伐他汀,用于测定中。
方法:测定用化合物处理的细胞或脑类器官中的DENV感染性和病毒体产生。各种方法,包括细胞和大脑类器官成像,进行蛋白质和基因检测以探索其抗病毒机制。
结果:结果揭示了Hernandonine的显着抗病毒特性,特别是在感染早期抑制DENV。机理分析表明,类似于genistein,Wortmannin,甲基-β-环糊精(MβCD),还有洛伐他汀,雌蕊对富含胆固醇的脂筏有影响。它还抑制了细胞的伪足运动能力,可能通过下调细胞骨架和内吞调节基因或蛋白质表达。此外,证明了雌草素的杀病毒活性。Hernandonine对DENV感染的抑制作用在疾病相关iPSC衍生的脑类器官模型中得到进一步验证,新型DENV-2感染系统值得进一步应用。
结论:这项研究证明了Hernandonine作为对抗DENV感染的新型候选药物的潜力。
BACKGROUND: Dengue, caused by the dengue virus (Orthoflavivirus dengue, encompassing DENV types 1-4), is a member of the Flaviviridae family. The symptoms of dengue range from subclinical or mild manifestations to potentially fatal complications. The management of severe dengue is exceptionally challenging due to the absence of effective antiviral medications. In this context, natural products, whether in the form of pure compounds or standardized plant extracts, have emerged as a promising source for the development of novel antiviral therapeutics. Hernandonine, an oxoaporphine alkaloid found in Hernandia nymphaeifolia (C. Presl) Kubitzki. serves both as a metabolite and an inhibitor of human immunodeficiency virus type 1 (HIV-1) integrase.
OBJECTIVE: This study investigated the ability of hernandonine to inhibit DENV infection and explored its potential mechanisms.
METHODS: To assess the in vitro anti-DENV activity, cells or induced pluripotent stem cell (iPSC)-derived cerebral organoids were exposed to hernandonine before or after infection with DENV. Along with hernandonine, the endocytosis modulators, genistein, wortmannin, Methyl-β-cyclodextrin (MβCD) and lovastatin, were used in the assays.
METHODS: The DENV infectivity and virion production in cells or cerebral organoids treated with compounds were determined. Various methods, including cell and cerebral organoids imaging, protein and gene detection were conducted to explore their antiviral mechanisms.
RESULTS: The results revealed notable antiviral properties of hernandonine, particularly in inhibiting DENV during the early stages of infection. Mechanistic analysis demonstrated that, akin to genistein, wortmannin, methyl-β-cyclodextrin (MβCD), and lovastatin, hernandonine exerted an influence on cholesterol-rich lipid rafts. It also restrained the pseudopodial movement ability of cells, potentially through the downregulation of cytoskeleton and endocytosis regulatory genes or protein expression. Moreover, hernandonine\'s virucidal activity was demonstrated. Hernandonine\'s inhibition of DENV infection was further validated in a disease-relevant iPSC-derived cerebral organoids model, a novel DENV-2 infection system worthy of further application.
CONCLUSIONS: This study evidenced the potential of hernandonine as a novel candidate in the fight against DENV infection.