Division of Chest Medicine, Department of Internal Medicine, Asia University Hospital, Taichung, Taiwan, ROC.
Division of Chest Medicine, Department of Internal Medicine, National Yang Ming Chiao Tung University Hospital, Taiwan, ROC; School of Medicine, College of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan, ROC.
J Microbiol Immunol Infect. 2023 Dec;56(6):1147-1157. doi: 10.1016/j.jmii.2023.09.003. Epub 2023 Sep 22.
SARS-CoV-2 spike proteins (SP) can bind to the human angiotensin-converting enzyme 2 (ACE2) in human pulmonary alveolar epithelial cells (HPAEpiC) and trigger an inflammatory process. Angiotensin-(1-7) may have an anti-inflammatory effect through activation of Mas receptor. This study aims to investigate whether SARS-CoV-2 SP can induce inflammation through ACE2 in the alveolar epithelial cells which can be modulated through angiotensin-(1-7)/Mas receptor axis.
HPAEpiC were treated with SARS-CoV-2 SP in the presence or absence of ACE2 antagonist-dalbavancin and Mas receptor agonist-angiotensin-(1-7). Proinflammatory cytokine production (IL-6 and IL-8) were measured at mRNA and protein levels. MAP kinase phosphorylation and transcription factor activation was determined by Western Blot. Mas receptor was blocked by either antagonist (A779) or knockdown (specific SiRNA). Experiments were replicated using A549 cells.
SARS-CoV-2 SP (5 μg/mL) significantly induced MAP kinase (ERK1/2) phosphorylation, downstream transcription factor (activator protein-1, AP-1) activation and cytokine production (IL-6 and IL-8) at both mRNA and protein levels. Pretreatment with dalbavancin (10 μg/mL), or angiotensin-(1-7) (10 μM) significantly reduced ERK1/2 phosphorylation, AP-1 activation, and cytokine production. However, these angiotensin-(1-7)-related protective effects were significantly abolished by blocking Mas receptor with either antagonist (A799,10 μM) or SiRNA knockdown.
SARS-CoV-2 SP can induce proinflammatory cytokine production, which can be inhibited by either ACE2 antagonist or Mas receptor agonist-angiotensin-(1-7). Angiotensin-(1-7)-related protective effect on cytokine reduction can be abolished by blocking Mas receptor. Our findings suggest that ACE2/angiotensin-(1-7)/Mas axis may serve as a therapeutic target to control inflammatory response triggered by SARS-CoV-2 SP.
SARS-CoV-2 刺突蛋白（SP）可与人类肺泡上皮细胞（HPAEpiC）中的人血管紧张素转换酶 2（ACE2）结合，并引发炎症过程。血管紧张素-(1-7)可通过激活 Mas 受体发挥抗炎作用。本研究旨在探讨 SARS-CoV-2 SP 是否可通过肺泡上皮细胞中的 ACE2 诱导炎症，而这种炎症可通过血管紧张素-(1-7)/Mas 受体轴来调节。
用 SARS-CoV-2 SP 处理 HPAEpiC，同时存在或不存在 ACE2 拮抗剂-达巴万星和 Mas 受体激动剂-血管紧张素-(1-7)。通过实时 PCR 和 Western blot 检测促炎细胞因子（IL-6 和 IL-8）的产生。通过 Western blot 检测 MAP 激酶磷酸化和转录因子激活。Mas 受体通过拮抗剂（A779）或 siRNA 敲低来阻断。实验使用 A549 细胞重复进行。
SARS-CoV-2 SP（5 μg/mL）可显著诱导 MAP 激酶（ERK1/2）磷酸化、下游转录因子（激活蛋白-1，AP-1）激活和细胞因子（IL-6 和 IL-8）的 mRNA 和蛋白水平表达。用达巴万星（10 μg/mL）或血管紧张素-(1-7)（10 μM）预处理可显著降低 ERK1/2 磷酸化、AP-1 激活和细胞因子产生。然而，用拮抗剂（A799，10 μM）或 siRNA 敲低阻断 Mas 受体可显著消除这些血管紧张素-(1-7)相关的保护作用。
SARS-CoV-2 SP 可诱导促炎细胞因子的产生，该产生可被 ACE2 拮抗剂或 Mas 受体激动剂-血管紧张素-(1-7)抑制。阻断 Mas 受体可消除血管紧张素-(1-7)对细胞因子减少的保护作用。我们的发现表明，ACE2/血管紧张素-(1-7)/Mas 轴可能成为控制 SARS-CoV-2 SP 触发的炎症反应的治疗靶点。
