Laboratory for Experimental Cardiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands.
Wilhelmina Children's Hospital Birth Center, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands.
Cells. 2020 Feb 18;9(2):468. doi: 10.3390/cells9020468.
Introduction: Preeclampsia (PE) represents a hypertensive pregnancy disorder that is associated with increased cardiovascular disease (CVD) risk. This increased risk has been attributed to accelerated atherosclerosis, with inflammation being a major contributor. Neutrophils play an important role in the onset and progression of atherosclerosis and have been associated with vascular damage in the placenta as well as the chronic inflammatory state in women with PE. We therefore investigated whether circulating neutrophil numbers or reactivity were associated with the presence and severity of subclinical atherosclerosis in women with a history of PE.
Women aged 45-60 years with a 10 to 20 years earlier history of early onset preeclampsia (delivery <34 weeks of gestation) (n = 90), but without symptomatic CVD burden were screened for the presence of subclinical coronary artery disease (CAD) using both contrast-enhanced and non-contrast coronary CT angiography. Subclinical CAD was defined as a coronary artery calcium (CAC) score ≥100 Agatston Units and/or ≥50% coronary luminal stenosis. We assessed whether the numbers and activity of circulating neutrophils were associated with the presence of subclinical CAD and as secondary outcome measurements, with the presence of any calcium (CAC score > 0 AU) or stenosis, categorized as absent (0%), minimal to mild (>0 and <50%), and moderate to severe (≥50%) narrowing of the coronary artery. Blood was drawn just before CT and neutrophil numbers were assessed by flow cytometry. In addition, the presence of the chemokine receptors CXCR2 and CXCR4, which are known to be instrumental in neutrophil recruitment, and neutrophil activity upon stimulation with the bacterial peptide N-Formylmethionyl-leucyl-phenylalanine (fMLF) was assessed by flow cytometry.
Of the participating women, with an average age of 49 years, 13% (12 out of 90) presented with subclinical signs of CAD (CAC score ≥100 AU and/or ≥50% luminal stenosis), and 37% (33 out of 90) had a positive CAC score (>0). Total white blood cell count and neutrophil counts were not associated with the presence of subclinical CAD or with a positive CAC score. When assessing the presence of the chemokine receptors CXCR4 and CXCR2, we observed a slight decrease of neutrophil CXCR2 expression in women with CAC (median MFI 22.0 [interquartile range (IQR) 20.2-23.8]) compared to women without CAC (23.8 [IQR 21.6-25.6], p = 0.02). We observed no differences regarding neutrophil CXCR4 expression. In addition, expression of the early activity marker CD35 was slightly lower on neutrophils of women with subclinical CAD (median MFI 1.6 [IQR 1.5-1.9] compared to 1.9 [IQR 1.7-2.1] in women without CAD, p = 0.02). However, for all findings, statistical significance disappeared after adjustment for multiple testing.
Our findings indicate that neutrophil counts and (re)activity are not directly associated with silent CAD disease burden and as such are not suitable as biomarkers to predict the presence of subclinical CAD in a high-risk population of women with a history of preeclampsia.
介绍：子痫前期（PE）是一种高血压妊娠疾病，与心血管疾病（CVD）风险增加有关。这种风险增加归因于动脉粥样硬化的加速，其中炎症是主要贡献者。中性粒细胞在动脉粥样硬化的发生和进展中起重要作用，并与胎盘血管损伤以及患有 PE 的女性的慢性炎症状态有关。因此，我们研究了循环中性粒细胞数量或反应性是否与患有 PE 的女性亚临床动脉粥样硬化的存在和严重程度相关。
我们筛选了 90 名年龄在 45-60 岁之间、有 10-20 年前早发型子痫前期（分娩<34 周）病史的女性，这些女性没有症状性 CVD 负担，使用对比增强和非对比冠状动脉 CT 血管造影检查亚临床冠状动脉疾病（CAD）的存在。亚临床 CAD 定义为冠状动脉钙（CAC）评分≥100 个 Agatston 单位和/或≥50%的冠状动脉管腔狭窄。我们评估了循环中性粒细胞的数量和活性是否与亚临床 CAD 的存在相关，并作为次要测量结果，评估任何钙（CAC 评分>0 AU）或狭窄的存在，分为无（0%）、最小至轻度（>0 且<50%）和中度至重度（≥50%）狭窄。在 CT 检查前采血，通过流式细胞术评估中性粒细胞数量。此外，通过流式细胞术评估已知在中性粒细胞募集中起重要作用的趋化因子受体 CXCR2 和 CXCR4 的存在，以及细菌肽 N-甲酰基甲硫氨酸-亮氨酸-苯丙氨酸（fMLF）刺激下中性粒细胞的活性。
在参与研究的女性中，平均年龄为 49 岁，13%（90 名女性中的 12 名）有亚临床 CAD 迹象（CAC 评分≥100 AU 和/或≥50%管腔狭窄），37%（90 名女性中的 33 名）有阳性 CAC 评分（>0）。总白细胞计数和中性粒细胞计数与亚临床 CAD 或阳性 CAC 评分无关。当评估趋化因子受体 CXCR4 和 CXCR2 的存在时，我们观察到 CAC 女性中性粒细胞 CXCR2 表达略有下降（中位数 MFI 22.0[四分位距（IQR）20.2-23.8]）与无 CAC 的女性相比（23.8[IQR 21.6-25.6]，p=0.02）。我们观察到中性粒细胞 CXCR4 表达没有差异。此外，亚临床 CAD 女性中性粒细胞早期活性标志物 CD35 的表达略低（中位数 MFI 1.6[IQR 1.5-1.9]与无 CAD 的女性相比（1.9[IQR 1.7-2.1]，p=0.02）。然而，对于所有发现，在进行多次测试调整后，统计学意义消失。
我们的研究结果表明，中性粒细胞计数和（再）活性与无症状 CAD 疾病负担没有直接关系，因此不适用于预测患有子痫前期病史的高危人群中亚临床 CAD 的存在。
