Department of Anesthesia, Kyoto University Hospital, 54 Shogoin-kawahara-cho, Sakyo-Ku, Kyoto, 606-8507, Japan.
Department of Anesthesia, Kitano Hospital, 2-4-20 Ohgimachi, Kita-ku, Osaka, 530-8480, Japan.
Clin Exp Med. 2023 Nov;23(7):3701-3708. doi: 10.1007/s10238-023-01091-4. Epub 2023 May 19.
Thrombosis has been associated with severity and mortality in COVID-19. SARS-CoV-2 infects the host via its spike protein. However, direct effects of spike proteins from SARS-CoV-2 variants on platelet activity and coagulability have not been examined. An ethically approved ex vivo study was performed under a preplanned power analysis. Venous blood was collected from 6 healthy subjects who gave prior written consent. The samples were divided into 5 groups: without spike proteins (group N) and with spike proteins derived from alpha, beta, gamma, and delta SARS-CoV-2 variants (groups A, B, C, and D, respectively). Platelet aggregability, P-selectin expression, platelet-associated complement-1 (PAC-1) binding, platelet count, and mean platelet volume (MPV) were measured in all 5 groups, and thromboelastography (TEG) parameters were measured in groups N and D. The % change in each parameter in groups A to D was calculated relative to the value in group N. Data were analyzed by Friedman test, except for TEG parameters, which were evaluated by Wilcoxon matched pairs test. P < 0.05 was considered significant. This study included 6 participants based on a power analysis. There were no significant differences in platelet aggregability under stimulation with adenosine diphosphate 5 µg/ml, collagen 0.2 or 0.5 µg/ml, and Ser-Phe-Leu-Leu-Arg-Asn-amide trifluoroacetate salt (SFLLRN) 0.5 or 1 µM in groups A-D compared to group N. There were also no significant differences in P-selectin expression and PAC-1 binding under basal conditions or SFLLRN stimulation, and no significant differences in platelet count, MPV and TEG parameters. Platelet hyperactivity and blood hypercoagulability have been reported in COVID-19 patients, but spike proteins at 5 µg/ml from SARS-CoV-2 variants (alpha, beta, gamma, delta) did not directly cause these effects in an ex vivo study. This study was approved by the Ethics Committee of Kyoto University Hospital (R0978-1) on March 06, 2020.
血栓形成与 COVID-19 的严重程度和死亡率有关。SARS-CoV-2 通过其刺突蛋白感染宿主。然而，SARS-CoV-2 变体的刺突蛋白对血小板活性和可凝性的直接影响尚未被研究。根据预先计划的功率分析，进行了一项伦理批准的离体研究。从 6 名事先书面同意的健康受试者采集静脉血。将样本分为 5 组：无刺突蛋白组（N 组）和分别来自 alpha、beta、gamma 和 delta SARS-CoV-2 变体的刺突蛋白组（A、B、C 和 D 组）。在 5 组中均测量血小板聚集率、P-选择素表达、血小板相关补体 1（PAC-1）结合、血小板计数和平均血小板体积（MPV），并在 N 组和 D 组中测量血栓弹力图（TEG）参数。在 A 至 D 组中，各参数的%变化相对于 N 组的值计算。除 TEG 参数外，数据通过 Friedman 检验进行分析，而 TEG 参数通过 Wilcoxon 配对检验进行评估。认为 P<0.05 有统计学意义。根据功率分析，这项研究包括 6 名参与者。与 N 组相比，在 5μg/ml 二磷酸腺苷、0.2 或 0.5μg/ml 胶原蛋白和 0.5 或 1μM Ser-Phe-Leu-Leu-Arg-Asn-amide trifluoroacetate salt（SFLLRN）刺激下，A-D 组的血小板聚集率无显著差异。在基础条件或 SFLLRN 刺激下，P-选择素表达和 PAC-1 结合也无显著差异，血小板计数、MPV 和 TEG 参数也无显著差异。COVID-19 患者存在血小板活性亢进和血液高凝状态，但在离体研究中，SARS-CoV-2 变体（alpha、beta、gamma、delta）的 5μg/ml 刺突蛋白并未直接引起这些作用。这项研究于 2020 年 3 月 6 日获得京都大学医院伦理委员会的批准（R0978-1）。
