Small interfering RNA targeting for infected-cell polypeptide 4 inhibits herpes simplex virus type 1 replication in retinal pigment epithelial cells
Small interfering RNA targeting for infected-cell polypeptide 4 inhibits herpes simplex virus type 1 replication in retinal pigment epithelial cells
复制标题
靶向感染细胞多肽 4 的小干扰 RNA 抑制视网膜色素上皮细胞中 1 型单纯疱疹病毒复制
DOI：
10.1111/j.1442-9071.2011.02668.x
复制
发表时间：
2012-03-01
影响因子：
4
通讯作者：
Wu, Kaili
中科院分区：
文献类型：
--
作者：
Duan, Fang;Ni, Shiming;Wu, Kaili
背景资料：本研究旨在利用针对感染细胞多肽4基因的小分子干扰RNA（siRNA）介导单纯疱疹病毒1型裂解性（生产性）感染视网膜上皮细胞中早期和晚期病毒基因的转录，抑制单纯疱疹病毒1型复制。采用Western blot、空斑试验、间接免疫荧光和逆转录聚合酶链反应等方法评价小干扰RNA的抗病毒作用。结果：小干扰RNA转染前、后均可降低单纯疱疹病毒1型感染视网膜上皮细胞中感染细胞多肽4的表达。与单纯疱疹病毒1型感染或转染阴性对照小干扰RNA相比，转染感染细胞多肽4靶向小干扰RNA（50和100 nM）的视网膜上皮细胞中病毒滴度和视网膜上皮细胞病变效应显著降低（P < 0.05）。小干扰RNA能有效地抑制单纯疱疹病毒1型感染细胞多肽4在mRNA和蛋白水平的表达（P < 0.05）。Westernblot检测结果表明，靶向感染细胞多肽4的小干扰RNA对单纯疱疹病毒1型感染的人角膜上皮细胞、人眼小梁细胞和Vero细胞中感染细胞多肽4蛋白的表达有抑制作用。靶向视网膜细胞多肽4的小干扰RNA可以抑制单纯疱疹病毒1型在视网膜上皮细胞中的复制，为RNA干扰作为抗病毒治疗的发展提供了基础。
Background: This study sought to inhibit herpes simplex virus type 1 replication using small interfering RNA which targeting infected-cell polypeptide 4 genes to mediate transcription of early and late viral genes in herpes simplex virus type 1 lytic (productive) infection in retina epithelial cells.Methods: After pre- or post-infecting with herpes simplex virus type 1, small interfering RNAs were transfected into retina epithelial cells. The antiviral effects of small interfering RNA were evaluated by Western blot, plaque assays, indirect immunofluorescence and reverse transcription polymerase chain reaction. The viral titre was detected by the 50% tissue culture infective dose method.Results: Small interfering RNA decreased infected-cell polypeptide 4 expression in retina epithelial cells that were infected with herpes simplex virus type 1 before or after small interferingRNAtransfection. Compared with herpes simplex virus type 1 infection alone or transfection with negative control small interfering RNA, the viral titre and the retina epithelial cell cytopathic effect were significantly decreased in retina epithelial cells transfected with infected-cell polypeptide 4-targeting small interfering RNA (50 and 100 nM) (P < 0.05). The small interfering RNA effectively silenced herpes simplex virus type 1 infected-cell polypeptide 4 expression on both mRNA and the protein levels (P < 0.05). The inhibition of infected-cell polypeptide 4-targeting small interfering RNA on infected-cell polypeptide 4 protein expression was also verified by Western blot in herpes simplex virus type 1 infected human cornea epithelial cell, human trabecular meshwork cells and Vero cells.Conclusions: Infected-cell polypeptide 4-targeting small interfering RNA can inhibit herpes simplex virus type 1 replication in retina epithelial cells, providing a foundation for development of RNA interference as an antiviral therapy.