Inhibition of Chikungunya Virus Replication by Harringtonine, a Novel Antiviral That Suppresses Viral Protein Expression
Inhibition of Chikungunya Virus Replication by Harringtonine, a Novel Antiviral That Suppresses Viral Protein Expression
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DOI：
10.1128/aac.01467-12
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发表时间：
2013-01-01
影响因子：
4.9
通讯作者：
Chu, Justin Jang Hann
中科院分区：
文献类型：
--
作者：
Kaur, Parveen;Thiruchelvan, Meerra;Chu, Justin Jang Hann
基孔肯雅病毒（CHIKV）是一种蚊子传播的病毒，在过去十年中再次成为重大的公共卫生威胁。自2005-2006年基孔肯雅热在印度洋留尼汪岛流行以来，已有40多个国家的数百万人受到感染。尽管如此，目前还没有针对基孔肯雅感染的抗病毒治疗。在这项研究中，开发了一种基于免疫荧光的筛选平台，以鉴定CHIKV感染的潜在抑制剂。使用高度纯化的天然产物化合物文库进行初步筛选，并将表现出>= 70%的CHIKV感染抑制的44种化合物鉴定为阳性命中。在这些中，选择四种用于剂量依赖性抑制测定以确认它们的抗CHIKV活性。三尖杉酯碱，三尖杉碱生物碱，显示出有效的抑制CHIKV感染（50%有效浓度[EC 50] = 0.24 μ M），细胞毒性最小，并选择阐明其抗病毒机制。添加时间研究、共处理测定和病毒基因组RNA的直接转染表明，三尖杉酯碱抑制病毒进入细胞后发生的CHIKV复制周期的早期阶段。此外，定量逆转录-PCR（qRT-PCR）和Western印迹分析表明三尖杉酯碱影响CHIKV RNA产生以及病毒蛋白表达。三尖杉酯碱对辛德毕斯病毒（一种相关甲病毒）的治疗表明，三尖杉酯碱可以抑制其他甲病毒。本研究首次提示三尖杉酯碱通过抑制CHIKV病毒蛋白合成发挥其抗病毒作用。
Chikungunya virus (CHIKV) is a mosquito-transmitted virus that has reemerged as a significant public health threat in the last decade. Since the 2005-2006 chikungunya fever epidemic in the Indian Ocean island of La Reunion, millions of people in more than 40 countries have been infected. Despite this, there is currently no antiviral treatment for chikungunya infection. In this study, an immunofluorescence-based screening platform was developed to identify potential inhibitors of CHIKV infection. A primary screen was performed using a highly purified natural product compound library, and 44 compounds exhibiting >= 70% inhibition of CHIKV infection were identified as positive hits. Among these, four were selected for dose-dependent inhibition assays to confirm their anti-CHIKV activity. Harringtonine, a cephalotaxine alkaloid, displayed potent inhibition of CHIKV infection (50% effective concentration [EC50] = 0.24 mu M) with minimal cytotoxicity and was selected for elucidation of its antiviral mechanism. Time-of-addition studies, cotreatment assays, and direct transfection of viral genomic RNA indicated that harringtonine inhibited an early stage of the CHIKV replication cycle which occurred after viral entry into cells. In addition, quantitative reverse transcription-PCR (qRT-PCR) and Western blot analyses indicated that harringtonine affects CHIKV RNA production as well as viral protein expression. Treatment of harringtonine against Sindbis virus, a related alphavirus, suggested that harringtonine could inhibit other alphaviruses. This study suggests for the first time that harringtonine exerts its antiviral effects by inhibiting CHIKV viral protein synthesis.