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菖蒲 A 从菖蒲根中抑制登革热病毒的增殖和感染。

Tatanan A from the Acorus calamus L. root inhibited dengue virus proliferation and infections.

机构信息

State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of New Drug Screening, Guangzhou Key Laboratory of Drug Research for Emerging Virus Prevention and Treatment, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, PR China.

Department of Pharmaceutical and Life Sciences, Jiujiang University, Jiujiang 332005, PR China.

出版信息

Phytomedicine. 2018 Mar 15;42:258-267. doi: 10.1016/j.phymed.2018.03.018.

Abstract

BACKGROUND

Acorus calamus l. (Acoraceae) is a well-known traditional Chinese medicinal plant, whose root are historically mainly used to treat neurodegenerative diseases, and for cholera treatment. This datum strongly indicates the antimicrobial activity of A. calamus.

PURPOSE

Our goal is to find the active constituents of A. calamus to treat dengue virus (DENV) infections, and to study the effects and mechanisms of these active substances.

METHODS

The root of A. calamus was extracted by ethanol. Mosquito larva C6/36 cells were used for DENV2 replication and transfection host. Mouse kidney fibroblast cells (BHK-21) were used as a host cell to study the infection ability of the virus. DENV2-induced cytopathic effect (CPE) and plaque assay were used to evaluate the inhibitory effect of A. calamus extracts on DENV2 infectivity inhibition. The levels of E and NS1 protein expression were measured by real-time PCR and western blot assays.

RESULTS

12 compounds were isolated from ethanol extract of A. calamus root, tatanan A showed the best anti-DENV ability among these 12 compounds, which significantly alleviated DENV2-induced CPE and cytotoxicity effects, with an EC of 3.9 µM. In addition, RNA replication assay further confirmed the antivirus ability of tatanan A. Time-addition assay showed that tatanan A affected the early stage of viral RNA replication, which in turn inhibited mRNA and protein levels of DENV2.

CONCLUSIONS

These results demonstrated the anti-DENV2 effect of tatanan A, in inhibiting DENV2 RNA replication and infections. In summary, tatanan A was found to be a novel natural DENV inhibitor and a potential candidate for the treatment of DENV infectious disease.

摘要

背景

菖蒲(天南星科)是一种广为人知的传统中药,其根茎历史上主要用于治疗神经退行性疾病和霍乱。这一数据强烈表明菖蒲具有抗菌活性。

目的

我们的目标是寻找菖蒲的活性成分来治疗登革热病毒(DENV)感染,并研究这些活性物质的作用和机制。

方法

采用乙醇提取菖蒲根茎。用蚊子幼虫 C6/36 细胞进行 DENV2 复制和转染宿主。用小鼠肾成纤维细胞(BHK-21)作为宿主细胞研究病毒的感染能力。用 DENV2 诱导的细胞病变效应(CPE)和蚀斑试验评估菖蒲提取物对 DENV2 感染性的抑制作用。用实时 PCR 和 Western blot 检测 E 和 NS1 蛋白表达水平。

结果

从菖蒲根茎的乙醇提取物中分离出 12 种化合物,其中 tatanan A 对 12 种化合物表现出最好的抗 DENV 能力,它能显著减轻 DENV2 诱导的 CPE 和细胞毒性作用,EC 为 3.9µM。此外,RNA 复制试验进一步证实了 tatanan A 的抗病毒能力。时间添加试验表明,tatanan A 影响病毒 RNA 复制的早期阶段,进而抑制 DENV2 的 mRNA 和蛋白水平。

结论

这些结果表明 tatanan A 具有抗 DENV2 作用,能抑制 DENV2 RNA 复制和感染。综上所述,tatanan A 被发现是一种新型天然 DENV 抑制剂,是治疗 DENV 感染性疾病的潜在候选药物。

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