Jinzhou Medical University, Jinzhou, Liaoning, China.
Shenzhen Eye Hospital, Jinan University, Shenzhen Eye Institute, Shenzhen, Guangdong, China.
Invest Ophthalmol Vis Sci. 2024 Jan 2;65(1):38. doi: 10.1167/iovs.65.1.38.
Whether H1N1 infection-associated ocular manifestations result from direct viral infections or systemic complications remains unclear. This study aimed to comprehensively elucidate the underlying causes and mechanism.
TCID50 assays was performed at 24, 48, and 72 hours to verify the infection of H1N1 in human retinal microvascular endothelial cells (HRMECs). The changes in gene expression profiles of HRMECs at 24, 48, and 72 hours were characterized using RNA sequencing technology. Differentially expressed genes (DEGs) were validated using real-time quantitative polymerase chain reaction and Western blotting. CCK-8 assay and scratch assay were performed to evaluate whether there was a potential improvement of proliferation and migration in H1N1-infected cells after oseltamivir intervention.
H1N1 can infect and replicate within HRMECs, leading to cell rounding and detachment. After H1N1 infection of HRMECs, 2562 DEGs were identified, including 1748 upregulated ones and 814 downregulated ones. These DEGs primarily involved in processes such as inflammation and immune response, cytokine-cytokine receptor interaction, signal transduction regulation, and cell adhesion. The elevated expression levels of CXCL10, CXCL11, CCL5, TLR3, C3, IFNB1, IFNG, STAT1, HLA, and TNFSF10 after H1N1 infection were reduced by oseltamivir intervention, reaching levels comparable to those in the uninfected group. The impaired cell proliferation and migration after H1N1 infection was improved by oseltamivir intervention.
This study confirmed that H1N1 can infect HRMECs, leading to the upregulation of chemokines, which may cause inflammation and destruction of the blood-retina barrier. Moreover, early oseltamivir administration may reduce retinal inflammation and hemorrhage in patients infected with H1N1.
甲型 H1N1 感染相关的眼部表现是由病毒直接感染还是全身并发症引起尚不清楚。本研究旨在全面阐明其潜在病因和机制。
在 24、48 和 72 小时进行 TCID50 测定，以验证甲型 H1N1 在人视网膜微血管内皮细胞（HRMECs）中的感染情况。采用 RNA 测序技术分析 HRMECs 在 24、48 和 72 小时的基因表达谱变化。采用实时定量聚合酶链反应和 Western blot 验证差异表达基因（DEGs）。CCK-8 检测和划痕实验评估奥司他韦干预后 H1N1 感染细胞的增殖和迁移是否有潜在改善。
甲型 H1N1 可在 HRMECs 内感染和复制，导致细胞圆化和脱落。HRMECs 感染甲型 H1N1 后，鉴定出 2562 个差异表达基因，其中上调基因 1748 个，下调基因 814 个。这些 DEGs 主要参与炎症和免疫反应、细胞因子-细胞因子受体相互作用、信号转导调节和细胞黏附等过程。甲型 H1N1 感染后，CXCL10、CXCL11、CCL5、TLR3、C3、IFNB1、IFNG、STAT1、HLA 和 TNFSF10 的表达水平升高，奥司他韦干预后降低，与未感染组相当。奥司他韦干预后可改善 H1N1 感染后细胞增殖和迁移受损。
本研究证实甲型 H1N1 可感染 HRMECs，导致趋化因子上调，可能引起炎症和血视网膜屏障破坏。此外，早期奥司他韦给药可能减轻甲型 H1N1 感染患者的视网膜炎症和出血。
