Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul National University Bundang Hospital, Seongnam, Korea.
Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul National University Boramae Hospital, Seoul, Korea.
PLoS One. 2020 Dec 31;15(12):e0244720. doi: 10.1371/journal.pone.0244720. eCollection 2020.
We sought to identify novel biomarkers in the amniotic fluid (AF) related to imminent spontaneous preterm delivery (SPTD) (≤ 14 days after sampling) in women with early preterm premature rupture of membranes (PPROM), using a protein microarray.
This was a retrospective cohort study of a total of 88 singleton pregnant women with PPROM (23+0 to 30+6 weeks) who underwent amniocentesis. A nested case-control study for biomarker discovery was conducted using pooled AF samples from controls (non-imminent delivery, n = 15) and cases (imminent SPTD, n = 15), which were analyzed using an antibody microarray. Quantitative validation of four candidate proteins was performed, using ELISA, in the total cohort (n = 88). IL-8, MMP-9, and Fas levels were additionally measured for the comparison and to examine association of SPTD with the etiologic factors of PPROM.
Of all the proteins studied in the protein microarray, four showed significant intergroup differences. Analyses of the total cohort by ELISA confirmed the significantly elevated concentrations of AF lipocalin-2, MMP-9, and S100 A8/A9, but not of endostatin and Fas, in women who delivered within 14 days of sampling. For inflammatory proteins showing a significant association, the odds of SPTD within 14 days increased significantly with an increase in baseline AF levels of the proteins (P for trend <0.05 for each) in each quartile, especially in the 3rd and 4th quartile.
We identified several potential novel biomarkers (i.e., lipocalin-2, MMP-9, and S100 A8/A9) related to SPTD within 14 days of sampling, all of which are inflammation-related molecules. Furthermore, the SPTD risk increased with increasing quartiles of each of these inflammatory proteins, especially the 3rd and 4th quartile of each protein. The present findings may highlight the importance of inflammatory mechanisms and the degree of activated inflammatory response in developing SPTD in early PPROM.
本研究旨在通过蛋白质微阵列，鉴定与早发性胎膜早破（PPROM）孕妇发生即将发生的自发性早产（SPTD）（≤ 采样后 14 天）相关的羊水（AF）中新型生物标志物。
这是一项回顾性队列研究，共纳入 88 例早发性 PPROM（23+0 至 30+6 周）孕妇，行羊膜穿刺术。使用来自对照组（非即将分娩，n=15）和病例组（即将发生的 SPTD，n=15）的混合 AF 样本进行生物标志物发现的嵌套病例对照研究，并用抗体微阵列进行分析。使用 ELISA 对总队列（n=88）中的 4 种候选蛋白进行定量验证。另外，还测量了 IL-8、MMP-9 和 Fas 水平，以比较 SPTD 与 PPROM 的病因因素的关系。
在蛋白质微阵列研究的所有蛋白中，有 4 种蛋白显示出显著的组间差异。ELISA 分析总队列的结果证实，在采样后 14 天内分娩的妇女的 AF 脂联素-2、MMP-9 和 S100A8/A9 浓度明显升高，但内抑素和 Fas 浓度没有升高。对于与 SPTD 显著相关的炎症蛋白，在每个四分位数中，随着蛋白基线 AF 水平的增加（每个四分位数 P 趋势值均<0.05），14 天内发生 SPTD 的可能性显著增加，尤其是在第 3 四分位数和第 4 四分位数中。
我们鉴定了几种与采样后 14 天内 SPTD 相关的潜在新型生物标志物（即脂联素-2、MMP-9 和 S100A8/A9），它们都是炎症相关分子。此外，这些炎症蛋白的每个四分位数的 SPTD 风险均增加，尤其是每个蛋白的第 3 四分位数和第 4 四分位数。这些发现可能突出了炎症机制和炎症反应激活程度在早发性 PPROM 中发生 SPTD 的重要性。
