活性氧与 N-(4-羟苯基)-视黄酰胺对严重急性呼吸综合征-冠状病毒 2 进入的抑制作用有关,The Journal of Biochemistry

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活性氧与 N-(4-羟苯基)-视黄酰胺对严重急性呼吸综合征-冠状病毒 2 进入的抑制作用有关
The Journal of Biochemistry ( IF 2.1 ) Pub Date : 2023-03-08 , DOI: 10.1093/jb/mvad020
Yasuhiro Hayashi ₁ , Xuhao Huang ₂ , Takashi Tanikawa ₃ , Kazunari Tanigawa ₄ , Mizuki Yamamoto ₅ , Jin Gohda ₅ , Jun-Ichiro Inoue ₆ , Koichi Fukase _{2,

7} , Kazuya Kabayama _{2,

7}

Affiliation

N-(4-hydroxyphenyl)-retinamide (4-HPR) 抑制二氢神经酰胺 Δ4-去饱和酶 1 (DEGS1) 酶活性。我们之前报道过，4-HPR 通过以不依赖于 DEGS1 的方式降低膜流动性来抑制严重急性呼吸系统综合症冠状病毒 2 (SARS-CoV-2) 刺突蛋白介导的膜融合。然而，4-HPR 抑制病毒进入的确切机制仍不清楚。在这项研究中，我们检查了活性氧 (ROS) 在 4-HPR 抑制膜融合中的作用，因为 4-HPR 是众所周知的 ROS 诱导剂。在 4-HPR 处理后的细胞-细胞融合测定中，发现靶细胞中细胞内 ROS 的产生增加，这通过添加抗氧化剂 α-生育酚 (TCP) 减弱。通过添加 TCP 减轻了细胞-细胞融合试验中 4-HPR 处理对膜融合敏感性的降低。此外，光漂白分析后的荧光恢复表明，糖基磷脂酰肌醇锚定蛋白和 SARS CoV-2 受体的横向扩散通过 4-HPR 处理减少，并通过添加 TCP 恢复。这些结果表明，4-HPR 降低 SARS-CoV-2 刺突蛋白介导的膜融合和膜流动性是由于 ROS 的产生。总而言之，这些结果表明 ROS 的产生与 4-HPR 对 SARS-CoV-2 进入的抑制作用有关。光漂白分析后的荧光恢复表明，糖基磷脂酰肌醇锚定蛋白和 SARS CoV-2 受体的横向扩散通过 4-HPR 处理减少，并通过添加 TCP 恢复。这些结果表明，4-HPR 降低 SARS-CoV-2 刺突蛋白介导的膜融合和膜流动性是由于 ROS 的产生。总而言之，这些结果表明 ROS 的产生与 4-HPR 对 SARS-CoV-2 进入的抑制作用有关。光漂白分析后的荧光恢复表明，糖基磷脂酰肌醇锚定蛋白和 SARS CoV-2 受体的横向扩散通过 4-HPR 处理减少，并通过添加 TCP 恢复。这些结果表明，4-HPR 降低 SARS-CoV-2 刺突蛋白介导的膜融合和膜流动性是由于 ROS 的产生。总而言之，这些结果表明 ROS 的产生与 4-HPR 对 SARS-CoV-2 进入的抑制作用有关。

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Reactive oxygen species are associated with the inhibitory effect of N-(4-hydroxyphenyl)-retinamide on the entry of the severe acute respiratory syndrome-coronavirus 2

N-(4-hydroxyphenyl)-retinamide (4-HPR) inhibits the dihydroceramide Δ4-desaturase 1 (DEGS1) enzymatic activity. We previously reported that 4-HPR suppresses the severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) spike protein-mediated membrane fusion through a decrease in membrane fluidity in a DEGS1-independent manner. However, the precise mechanism underlying the inhibition of viral entry by 4-HPR remains unclear. In this study, we examined the role of reactive oxygen species (ROS) in the inhibition of membrane fusion by 4-HPR, since 4-HPR is a well-known ROS-inducing agent. Intracellular ROS generation was found to be increased in the target cells in a cell–cell fusion assay following 4-HPR treatment, which was attenuated by the addition of the antioxidant, α-tocopherol (TCP). The reduction in membrane fusion susceptibility by 4-HPR treatment in the cell–cell fusion assay was alleviated by TCP addition. Furthermore, fluorescence recovery after photobleaching analysis showed that the lateral diffusion of glycosylphosphatidylinositol-anchored protein and SARS CoV-2 receptor was reduced by 4-HPR treatment and restored by TCP addition. These results indicate that the decrease in SARS-CoV-2 spike protein-mediated membrane fusion and membrane fluidity by 4-HPR was due to ROS generation. Taken together, these results demonstrate that ROS production is associated with the 4-HPR inhibitory effect on SARS-CoV-2 entry.

更新日期：2023-03-08

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