Guangzhou Institute of Cardiovascular Disease, The Second Affiliated Hospital, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, China; Department of Histology and Embryology, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, China.
Department of Histology and Embryology, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, China.
Exp Eye Res. 2019 Aug;185:107664. doi: 10.1016/j.exer.2019.05.004. Epub 2019 May 11.
HSV-1 infection in corneal epithelium initiates the process of herpes simplex keratitis. We investigated the dynamic change of the host proteins in corneal epithelial cells infected with HSV-1 to understand the virus-host interaction. iTRAQ coupled with LC-MS/MS was applied to quantitatively analyze the protein profiles in HSV-1 infected corneal epithelial cells at 6 and 24 h post-infection (hpi), and the results were validated by multiple reaction monitoring (MRM). We also performed bioinformatic analysis to investigate the potentially important signal pathways and protein interaction networks in the host response to HSV-1 infection. We identified 292 proteins were up-regulated and 168 proteins were down-regulated at 6 hpi, while 132 proteins were up-regulated and 89 proteins were down-regulated at 24 hpi, which were validated by MRM analysis. We found the most enriched GO terms were translational initiation, cytosol, poly(A) RNA binding, mRNA splicing via spliceosome and extracellular exosome for the dysregulated proteins. KEGG pathway analysis revealed significant changes in metabolism pathway characterized by decreased tricarboxylic acid cycle activity and increased glycolysis. Proteins interaction network analysis indicated several proteins including P4HB, ACLY, HSP90AA1 and EIF4A3, might be critical proteins in the host-virus response. Our study for the first time analyzed the protein profile of HSV-1 infected primary corneal epithelial cells by quantitative proteomics. These findings help to better understand the host-virus interaction and the pathogenesis of herpes simplex keratitis.
单纯疱疹病毒 1（HSV-1）感染角膜上皮细胞，引发单纯疱疹病毒性角膜炎。我们研究了 HSV-1 感染角膜上皮细胞后宿主蛋白的动态变化，以了解病毒-宿主相互作用。我们采用 iTRAQ 联合 LC-MS/MS 技术，定量分析了 HSV-1 感染角膜上皮细胞后 6 和 24 小时（hpi）的蛋白谱，并通过多重反应监测（MRM）进行了验证。我们还进行了生物信息学分析，以研究宿主对 HSV-1 感染反应中的潜在重要信号通路和蛋白相互作用网络。我们鉴定出 292 个蛋白在 6 hpi 时上调，168 个蛋白下调，而在 24 hpi 时，132 个蛋白上调，89 个蛋白下调，这些结果通过 MRM 分析得到了验证。我们发现，最富集的 GO 术语是翻译起始、细胞质、聚（A）RNA 结合、通过剪接体进行 mRNA 剪接和细胞外外泌体，这些术语与失调蛋白相关。KEGG 通路分析显示，代谢途径发生了显著变化，三羧酸循环活性降低，糖酵解增加。蛋白相互作用网络分析表明，P4HB、ACLY、HSP90AA1 和 EIF4A3 等几种蛋白可能是宿主-病毒反应中的关键蛋白。本研究首次通过定量蛋白质组学分析了 HSV-1 感染原代角膜上皮细胞的蛋白谱。这些发现有助于更好地理解宿主-病毒相互作用和单纯疱疹病毒性角膜炎的发病机制。
