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Invest Ophthalmol Vis Sci. 2020 Aug 3;61(10):5. doi: 10.1167/iovs.61.10.5.
To examine the role of aqueous tumor necrosis factor α (TNF-α)-RhoA-Rho kinase (ROCK) signaling in cytomegalovirus (CMV)-induced apoptosis and the barrier function of cultured human corneal endothelial cells (hCECs) in CMV-positive Posner-Schlossman syndrome (CMV+/PSS) patients.
Aqueous levels of TNF-α, IL-8, IL-10, and several other cytokines in 19 CMV+/PSS patients and 20 healthy control subjects were quantitated using a multiplex assay. The expression of active RhoA in hCECs post-CMV infection was determined using western blotting (WB). The expression levels of TNF-α and nuclear factor kappa B (NF-κB) in CMV-infected hCECs were examined by immunocytochemistry (ICC) and WB with and without ROCK inhibitors. The apoptotic rate and barrier integrity in CMV-infected hCECs were also examined.
The expression levels of TNF-α, monocyte chemoattractant protein-1 (MCP-1), IL-8, and IL-10 were upregulated in the aqueous humor of CMV+/PSS patients, and among these upregulated cytokines aqueous TNF-α was negatively correlated with the number of corneal endothelial cells. In CMV-infected hCECs, upregulation of TNF-α and NF-κB was determined by WB and ICC. In hCECs, CMV infection induced apoptosis and significantly impaired cell-cell contacts, effects that were attenuated by treatment with a ROCK inhibitor.
Aqueous TNF-α was upregulated in CMV+/PSS patients, which may have triggered corneal endothelial cell loss. Modulation of TNF-α, including its downstream Rho-ROCK signaling, could serve as a novel treatment modality for corneal endothelial cell loss in CMV+/PSS patients.
研究细胞液中肿瘤坏死因子-α（TNF-α）-RhoA- Rho 激酶（ROCK）信号在巨细胞病毒（CMV）诱导的凋亡和 CMV 阳性 Posner-Schlossman 综合征（CMV+/PSS）患者培养的人角膜内皮细胞（hCEC）屏障功能中的作用。
使用多重分析检测 19 例 CMV+/PSS 患者和 20 例健康对照者的细胞液中 TNF-α、IL-8、IL-10 及其他几种细胞因子的水平。采用 Western blot（WB）检测 CMV 感染后 hCEC 中活性 RhoA 的表达。通过免疫细胞化学（ICC）和 WB 检测 CMV 感染的 hCEC 中 TNF-α和核因子 kappa B（NF-κB）的表达水平，并在有无 ROCK 抑制剂的情况下进行检测。还检测了 CMV 感染的 hCEC 的凋亡率和屏障完整性。
CMV+/PSS 患者的房水中 TNF-α、单核细胞趋化蛋白-1（MCP-1）、IL-8 和 IL-10 的表达水平上调，在这些上调的细胞因子中，细胞液 TNF-α与角膜内皮细胞数量呈负相关。通过 WB 和 ICC 确定了 CMV 感染 hCEC 中 TNF-α和 NF-κB 的上调。在 hCEC 中，CMV 感染诱导细胞凋亡，并显著破坏细胞间接触，用 ROCK 抑制剂处理可减轻这些作用。
CMV+/PSS 患者的房水中 TNF-α 上调，这可能引发了角膜内皮细胞的丧失。TNF-α的调节，包括其下游 Rho-ROCK 信号，可能成为 CMV+/PSS 患者角膜内皮细胞丧失的一种新的治疗方法。
