Department of Obstetrics and Gynecology, School of Medicine, Virginia Commonwealth University, Richmond, VA 23298-0034, USA.
Department of Physiology and Biophysics, School of Medicine, Virginia Commonwealth University, Richmond, VA 23298-0034, USA.
Int J Mol Sci. 2022 Oct 30;23(21):13218. doi: 10.3390/ijms232113218.
Neutrophils expressing cyclooxygenase-2 (COX-2) extensively infiltrate maternal blood vessels in preeclampsia, associated with vascular inflammation. Because pregnancy neutrophils also express protease-activated receptor 1 (PAR-1, F2R thrombin receptor), which they do not in non-pregnant subjects, they can be activated by proteases. We tested the hypothesis that aspirin at a dose sufficient to inhibit COX-2 would reduce inflammatory responses in preeclampsia neutrophils. Neutrophils were isolated from normal pregnant and preeclamptic women at approximately 30 weeks' gestation. Normal pregnancy neutrophils were treated with elastase, a protease elevated in preeclampsia, or elastase plus aspirin to inhibit COX-2, or elastase plus pinane thromboxane, a biologically active structural analog of thromboxane and a thromboxane synthase inhibitor. Preeclamptic pregnancy neutrophils were treated with the same doses of aspirin or pinane thromboxane. Confocal microscopy with immunofluorescence staining was used to determine the cellular localization of the p65 subunit of nuclear factor-kappa B (NF-κB) and media concentrations of thromboxane were measured to evaluate the inflammatory response. In untreated neutrophils of normal pregnant women, p65 was localized to the cytosol. Upon stimulation with elastase, p65 translocated from the cytosol to the nucleus coincident with increased thromboxane production. When neutrophils were co-treated with aspirin or pinane thromboxane, elastase was not able to cause nuclear translocation of p65 or increase thromboxane. In untreated neutrophils of preeclamptic women, the p65 subunit was present in the nucleus and thromboxane production was elevated, but when preeclamptic neutrophils were treated with aspirin or pinane thromboxane, p65 was cleared from the nucleus and returned to the cytosol along with decreased thromboxane production. These findings suggest that COX-2 is a downstream mediator of PAR-1 and demonstrate that PAR-1- mediated inflammation can be inhibited by aspirin. Given the extensive and ubiquitous expression of PAR-1 and COX-2 in preeclamptic women, consideration should be given to treating women with preeclampsia using a dose of aspirin sufficient to inhibit COX-2.
在子痫前期中，表达环氧化酶-2 (COX-2) 的中性粒细胞广泛浸润母体血管，与血管炎症有关。由于妊娠中性粒细胞也表达蛋白酶激活受体 1（PAR-1，凝血酶受体 F2R），而在非妊娠个体中则不表达，因此它们可以被蛋白酶激活。我们检验了这样一个假设，即阿司匹林的剂量足以抑制 COX-2，将减少子痫前期中性粒细胞的炎症反应。从大约 30 周妊娠的正常孕妇和子痫前期孕妇中分离中性粒细胞。用弹性蛋白酶（一种在子痫前期中升高的蛋白酶）或弹性蛋白酶加阿司匹林（抑制 COX-2）或弹性蛋白酶加 pinane 血栓烷（血栓烷的生物活性结构类似物和血栓烷合酶抑制剂）处理正常妊娠中性粒细胞。用相同剂量的阿司匹林或 pinane 血栓烷处理子痫前期妊娠中性粒细胞。用共聚焦显微镜免疫荧光染色来确定核因子-kappa B（NF-κB）的 p65 亚单位的细胞定位，并测量介质中的血栓烷浓度以评估炎症反应。在未经处理的正常妊娠妇女的中性粒细胞中，p65 定位于细胞质。用弹性蛋白酶刺激后，p65 从细胞质转移到细胞核，同时血栓烷的产生增加。当中性粒细胞同时用阿司匹林或 pinane 血栓烷处理时，弹性蛋白酶不能导致 p65 的核转位或增加血栓烷的产生。在未经处理的子痫前期妇女的中性粒细胞中，p65 亚单位存在于细胞核中，血栓烷的产生增加，但当用阿司匹林或 pinane 血栓烷处理子痫前期中性粒细胞时，p65 从细胞核中清除并返回细胞质，同时血栓烷的产生减少。这些发现表明 COX-2 是 PAR-1 的下游介质，并表明 PAR-1 介导的炎症可以被阿司匹林抑制。鉴于 PAR-1 和 COX-2 在子痫前期妇女中广泛且普遍的表达，应考虑使用足以抑制 COX-2 的阿司匹林剂量来治疗子痫前期妇女。
