Shi Jialing, Kuang Lin, Qi Li, Li Ruoyu, Wu Yangfan
School of Integrated Chinese and Western Medicine, Hunan University of Chinese Medicine, Changsha, China.
Xiangxing College of Hunan University of Chinese Medicine, Changsha, China.
Front Microbiol. 2025 Mar 24;16:1560340. doi: 10.3389/fmicb.2025.1560340. eCollection 2025.
The objective of this study was to investigate the effect of acyclovir (ACV) on the TLR9 signaling pathway after human immortalized epidermal (HaCaT) cell infection with herpes simplex virus type 2 (HSV-2).
In this study, an cell model of HSV-2 infection was successfully constructed by infecting HaCaT with HSV-2 virus. In order to explore the antiviral mechanism of acyclovir (ACV), high-throughput transcriptome sequencing (RNA-seq) was used to analyze the genome-wide expression profiling of infected cells before and after ACV treatment, and to systematically compare the change characteristics of differentially expressed genes (DEGs). Based on the sequencing results, the study further focused on Toll-like receptor (TLR) 9 signaling, using quantitative real-time reverse transcriptase chain reaction (qRT-PCR) to quantitatively detect the effect of ACV intervention on the mRNA expression level of key molecules of TLR 9 signaling pathway in HSV-2 infected HaCaT cells.
A total of 896 significant changes in gene expression were identified by the transcriptome analysis, including 314 upregulated genes and 582 downregulated genes. GO enrichment analysis showed that the differentially expressed genes were mainly related to CC includes the ubiquitin ligase complex, mitochondrial protein-containing complex, DNA-binding transcription activator activity, exonuclease activity, catabolic process, nuclear-transcribed mRNA catabolic process nuclear-transcribed mRNA catabolic process; KEGG enrichment analysis showed that the differentially expressed genes were mainly related to Toll-like receptor signaling pathway, herpes simplex virus 1 infection, and TNF signaling pathway. The RT-PCR results were confirmed to be basically consistent with the sequencing results.
ACV altered the transcriptome level of HSV-2 infection in HaCaT cells. The RT-PCR results confirmed that ACV intervened in HSV-2 infection through the TLR9 signaling pathway.
本研究的目的是探讨阿昔洛韦（ACV）对人永生化表皮细胞（HaCaT）感染2型单纯疱疹病毒（HSV-2）后Toll样受体9（TLR9）信号通路的影响。
本研究通过用HSV-2病毒感染HaCaT细胞成功构建了HSV-2感染的细胞模型。为了探究阿昔洛韦（ACV）的抗病毒机制，采用高通量转录组测序（RNA-seq）分析ACV处理前后感染细胞的全基因组表达谱，并系统比较差异表达基因（DEGs）的变化特征。基于测序结果，本研究进一步聚焦于Toll样受体（TLR）9信号通路，采用定量实时逆转录聚合酶链反应（qRT-PCR）定量检测ACV干预对HSV-2感染的HaCaT细胞中TLR 9信号通路关键分子mRNA表达水平的影响。
转录组分析共鉴定出896个基因表达的显著变化，包括314个上调基因和582个下调基因。基因本体（GO）富集分析表明，差异表达基因主要与包含泛素连接酶复合体、含线粒体蛋白复合体、DNA结合转录激活因子活性、核酸外切酶活性、分解代谢过程、核转录mRNA分解代谢过程的细胞成分（CC）相关；京都基因与基因组百科全书（KEGG）富集分析表明，差异表达基因主要与Toll样受体信号通路、单纯疱疹病毒1感染和肿瘤坏死因子（TNF）信号通路相关。逆转录聚合酶链反应（RT-PCR）结果经证实与测序结果基本一致。
ACV改变了HaCaT细胞中HSV-2感染的转录组水平。RT-PCR结果证实ACV通过TLR9信号通路干预HSV-2感染。
