A purified spike (S) glycoprotein of severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) coronavirus was used to study its effects on THP-1 macrophages, peripheral blood mononuclear cells (PBMCs), and HUVEC cells. The S protein mediates the entry of SARS-CoV-2 into cells through binding to the angiotensin-converting enzyme 2 (ACE2) receptors. We measured the viability, intracellular cytokine release, oxidative stress, proinflammatory markers, and THP-1-like macrophage polarization. We observed an increase in apoptosis, ROS generation, MCP-1, and intracellular calcium expression in the THP-1 macrophages. Stimulation with the S protein polarizes the THP-1 macrophages towards proinflammatory futures with an increase in the TNFα and MHC-II M1-like phenotype markers. Treating the cells with an ACE inhibitor, perindopril, at 100 µM reduced apoptosis, ROS, and MHC-II expression induced by S protein. We analyzed the sensitivity of the HUVEC cells after the exposure to a conditioned media (CM) of THP-1 macrophages stimulated with the S protein. The CM induced endothelial cell apoptosis and MCP-1 expression. Treatment with perindopril reduced these effects. However, the direct stimulation of the HUVEC cells with the S protein, slightly increased HIF1α and MCP-1 expression, which was significantly increased by the ACE inhibitor treatment. The S protein stimulation induced ROS generation and changed the mitogenic responses of the PBMCs through the upregulation of TNFα and interleukin (IL)-17 cytokine expression. These effects were reduced by the perindopril (100 µM) treatment. Proteomic analysis of the S protein stimulated THP-1 macrophages with or without perindopril (100 µM) exposed more than 400 differentially regulated proteins. Our results provide a mechanistic analysis suggesting that the blood and vascular components could be activated directly through S protein systemically present in the circulation and that the activation of the local renin angiotensin system may be partially involved in this process.

严重急性呼吸系统综合症相关冠状病毒 2 (SARS) 的纯化尖峰 (S) 糖蛋白——冠状病毒——2）冠状病毒用于研究其对THP-1巨噬细胞、外周血单核细胞（PBMC）和HUVEC细胞的影响。S 蛋白通过与血管紧张素转换酶 2 (ACE2) 受体结合，介导 SARS-CoV-2 进入细胞。我们测量了活力、细胞内细胞因子释放、氧化应激、促炎标志物和 THP-1 样巨噬细胞极化。我们观察到 THP-1 巨噬细胞中细胞凋亡、ROS 生成、MCP-1 和细胞内钙表达的增加。S 蛋白的刺激使 THP-1 巨噬细胞向促炎未来极化，TNFα 和 MHC-II M1 样表型标志物增加。用 100 µM 的 ACE 抑制剂培哚普利处理细胞可减少 S 蛋白诱导的细胞凋亡、ROS 和 MHC-II 表达。我们分析了 HUVEC 细胞在暴露于用 S 蛋白刺激的 THP-1 巨噬细胞的条件培养基 (CM) 后的敏感性。CM 诱导内皮细胞凋亡和 MCP-1 表达。培哚普利治疗减少了这些影响。然而，用 S 蛋白直接刺激 HUVEC 细胞后，HIF1α 和 MCP-1 的表达略微增加，ACE 抑制剂处理显着增加。S 蛋白刺激诱导 ROS 生成并通过上调 TNFα 和白细胞介素 (IL)-17 细胞因子表达改变 PBMC 的促有丝分裂反应。培哚普利 (100 µM) 处理降低了这些影响。S 蛋白的蛋白质组学分析刺激了 THP-1 巨噬细胞，有或没有培哚普利 (100 µM) 暴露超过 400 种差异调节的蛋白质。