天然罗汉松二萜体外抗登革病毒活性研究  

Anti-dengue virus activity of natural podocarpane-type diterpenoid in vitro

作　　者：汪芳 严欢[3] 姚债文 罗荣华[1] 刘海洋[3] 郑永唐[1] WANG Fang;YAN Huan;YAO Zhai-wen;LUO Rong-hua;LIU Hai-yang;ZHENG Yong-tang(Key Laboratory of Animal Models and Human Disease Mechanisms of Chinese Academy of Sciences,Kunming Institute of Zoology,Chinese Academy of Sciences,Kunming 650223,China;College of Medicine,Kunming University of Science and Technology,Kunming 650500,China;State Key Laboratory of Phytochemistry and Plant Resources in West China,Kunming Institute of Botany Chinese Academy of Sciences,Kunming 650201,China;College of Pharmaceutical Sciences,Soochow University,Suzhou 215021,China)

机构地区：[1]中国科学院昆明动物研究所,中国科学院动物模型与人类疾病机理重点实验室,云南昆明650223 [2]昆明理工大学医学院,云南昆明650500 [3]中国科学院昆明植物研究所,植物化学与西部植物资源持续利用国家重点实验室,云南昆明650201 [4]苏州大学药学院,江苏苏州215021

出　　处：《药学学报》2021年第3期793-798,共6页Acta Pharmaceutica Sinica

基　　金：云南省重大科技专项计划课题(2017ZF007)。

摘　　要：登革病毒(dengue virus,DENV)是造成热带和亚热带地区登革热和登革出血热季节性大爆发的蚊媒病原体,可导致严重威胁生命的疾病,因此亟需研发DENV疫苗和药物。本研究发现从石栗枝叶中分离的罗汉松型二萜(3α,5β,10α)-13-methoxypodocarpa-8,11,13-triene-3,12-diol(MPTD)具有很强的体外抗DENV作用。噬斑抑制实验检测MPTD对4种不同血清型DENV的抑制作用;MTT实验检测其对Vero和Huh7两种细胞的毒性;qRT-PCR和Western blot实验分别在RNA和蛋白水平检测其抗DENV的活性。结果表明,MPTD处理可显著降低DENV感染Vero细胞的病毒滴度,其对4种DENV(1~4)的半数有效浓度(50%effective concentration,EC50)值分别为2.72±0.39、10.99±5.18、18.72±0.21和0.48±0.28μmol·L^(-1)。MPTD显著抑制DENV RNA的合成和E蛋白的表达,其可能抑制DENV复制的前期阶段而发挥抗病毒活性。进一步的研究显示,MPTD对DENV感染的抑制作用不是靶向病毒进入细胞阶段。MPTD对DENV具有显著抑制作用,是一种有潜在应用价值的抗DENV化合物。Dengue virus(DENV)is the most rapidly transmitted mosquito-borne pathogen,which is the main cause of seasonal outbreaks of dengue fever and dengue hemorrhagic fever in tropical and subtropical regions,and may cause serious life-threatening diseases.There is an urgent need to develop effective vaccines or antiviral therapies.In this paper,we found that a podocarpane-type diterpenoid,(3α,5β,10α)-13-methoxypodocarpa-8,11,13-triene-3,12-diol(MPTD),isolated from the stems and leaves of Aleurites moluccana,showed good effect against DENV.The anti-DENV activity of MPTD against four different DENV serotypes was studied by plaque assay.The cytotoxicity of MPTD in Vero and Huh7 cells was tested by MTT assay.qRT-PCR and Western blot assays were used to investigate the anti-DENV activity of MPTD at RNA and protein levels,respectively.The results showed that MPTD greatly reduced the virus titer in DENV infected Vero cells,and its 50%effective concentration(EC50)for DENV(1–4)were 2.72±0.39,10.99±5.18,18.72±0.21,and 0.48±0.28μmol·L^(-1),respectively.The results showed that MPTD inhibits DENV RNA level and the expression of E protein.In addition,MPTD may inhibit the early stage of DENV replication and exert antiviral activity.Further studies showed that the inhibitory effect of MPTD against DENV infection is not targeting the viral entry stage.Therefore,MPTD has a significant anti-dengue virus effect,and is an anti-DENV compound with potential application value.

关 键 词：石栗 天然化合物 (3α 5β 10α)-13-methoxypodocarpa-8 11 13-triene-3 12-diol 罗汉松型二萜 登革病毒 抗病毒活性 

分 类 号：R966[医药卫生—药理学]