Dual Assay Validation of Rosmarinus officinalis Extract as an Inhibitor of SARS-CoV-2 Spike Protein: Combining Pseudovirus Testing, Yeast Two-Hybrid, and UPLC-Q Exactive Orbitrap-MS Profiling.

IF 3 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Phytochemical Analysis Pub Date : 2024-11-13 DOI:10.1002/pca.3467
Yujing Huang, Rufeng Luo, Chenjing Tian, Duntao Zu, Jianni Yang, Wenlin Chen, Dingqiang Huang, Siyan Duan, Shunxin Yan, Yujia Yuan, Shengrong Li, Haibo Zhou, Fulong Lin, Qinghui He, Junxia Zheng
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Abstract

Introduction: This study evaluates the effectiveness of Traditional Chinese Medicine (TCM) extracts in blocking the interaction between the SARS-CoV-2 Spike protein and human ACE2 receptor, utilizing a dual-method approach to explore the antiviral potential of natural compounds.

Objectives: This work aims to evaluate the capability of TCM extracts in inhibiting the SARS-CoV-2 Spike protein and ACE2 receptor interaction using advanced biochemical assays.

Methods: A dual-method screening approach was utilized, beginning with a pseudovirus assay to assess the inhibition capabilities of TCM extracts in vitro, followed by a split-ubiquitin yeast two-hybrid (Y2H) system to validate interactions in live cells. Active compounds were characterized and quantified using UPLC-Q-Exactive-Orbitrap-MS.

Results: Among the 91 TCM extracts tested, Rosmarinus officinalis exhibited the most potent inhibition in both pseudovirus and Y2H assays, significantly reducing viral entry and disrupting the Spike-ACE2 interaction. Comprehensive chemical profiling via UPLC-Q-Exactive-Orbitrap-MS identified 132 compounds, including phenolics, flavonoids, and terpenoids.

Conclusion: This research validates the use of TCM extracts in viral inhibition strategies, demonstrating the utility of integrating traditional remedies with modern scientific approaches to discover new therapeutic agents.

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将迷迭香提取物作为 SARS-CoV-2 Spike 蛋白抑制剂的双重测定验证:结合假病毒测试、酵母双杂交和UPLC-Q Exactive Orbitrap-MS分析。
引言本研究评估了中药提取物在阻断SARS-CoV-2 Spike蛋白和人类ACE2受体之间相互作用的有效性,利用双重方法探索天然化合物的抗病毒潜力:本研究旨在利用先进的生化检测方法评估中药提取物抑制SARS-CoV-2 Spike蛋白和ACE2受体相互作用的能力:该研究采用了双方法筛选方法,首先利用假病毒试验评估中药提取物在体外的抑制能力,然后利用分裂泛素酵母双杂交(Y2H)系统验证其在活细胞中的相互作用。采用 UPLC-Q-Exactive-Orbitrap-MS 对活性化合物进行了表征和定量:结果:在测试的91种中药提取物中,迷迭香在伪病毒和Y2H试验中都表现出了最有效的抑制作用,能显著减少病毒的进入并破坏Spike-ACE2的相互作用。通过UPLC-Q-Exactive-Orbitrap-MS进行的综合化学分析确定了132种化合物,包括酚类、类黄酮和萜类化合物:这项研究验证了中药提取物在病毒抑制策略中的应用,证明了将传统疗法与现代科学方法相结合以发现新的治疗药物的实用性。
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来源期刊
Phytochemical Analysis
Phytochemical Analysis 生物-分析化学
CiteScore
6.00
自引率
6.10%
发文量
88
审稿时长
1.7 months
期刊介绍: Phytochemical Analysis is devoted to the publication of original articles concerning the development, improvement, validation and/or extension of application of analytical methodology in the plant sciences. The spectrum of coverage is broad, encompassing methods and techniques relevant to the detection (including bio-screening), extraction, separation, purification, identification and quantification of compounds in plant biochemistry, plant cellular and molecular biology, plant biotechnology, the food sciences, agriculture and horticulture. The Journal publishes papers describing significant novelty in the analysis of whole plants (including algae), plant cells, tissues and organs, plant-derived extracts and plant products (including those which have been partially or completely refined for use in the food, agrochemical, pharmaceutical and related industries). All forms of physical, chemical, biochemical, spectroscopic, radiometric, electrometric, chromatographic, metabolomic and chemometric investigations of plant products (monomeric species as well as polymeric molecules such as nucleic acids, proteins, lipids and carbohydrates) are included within the remit of the Journal. Papers dealing with novel methods relating to areas such as data handling/ data mining in plant sciences will also be welcomed.
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