Weingrill Rodrigo Barbano, Paladino Sandra Luft, Souza Matheus Leite Ramos, Pereira Eduardo Manoel, Marques Aldilane Lays Xavier, Silva Elaine Cristina Oliveira, da Silva Fonseca Eduardo Jorge, Ursulino Jeferson Santana, Aquino Thiago Mendonça, Bevilacqua Estela, Urschitz Johann, Silva Jean Carl, Borbely Alexandre Urban
Programa de Pós-Graduação em Saúde e Meio Ambiente, Universidade da Região de Joinville - UNIVILLE, Joinville, Brazil.
Institute for Biogenesis Research, John A. Burns School of Medicine, University of Hawai'i at Mānoa, Honolulu, HI, United States.
Front Physiol. 2021 Dec 14;12:767112. doi: 10.3389/fphys.2021.767112. eCollection 2021.
Hypertensive disorders of pregnancy are closely associated with prematurity, stillbirth, and maternal morbidity and mortality. The onset of hypertensive disorders of pregnancy (HDP) is generally noticed after the 20th week of gestation, limiting earlier intervention. The placenta is directly responsible for modulating local and systemic physiology by communicating using mechanisms such as the release of extracellular vesicles, especially exosomes. In this study, we postulated that an analysis of exosome-enriched maternal plasma could provide a more focused and applicable approach for diagnosing HDP earlier in pregnancy. Therefore, the peripheral blood plasma of 24 pregnant women (11 controls, 13 HDP) was collected between 20th and 24th gestational weeks and centrifuged for exosome enrichment. Exosome-enriched plasma samples were analyzed by Raman spectroscopy and by proton nuclear magnetic resonance metabolomics (H NMR). Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to analyze the Raman data, from the spectral region of 600-1,800 cm, to determine its potential to discriminate between groups. Using principal component analysis, we were able to differentiate the two groups, with 89% of all variances found in the first three principal components. In patients with HDP, most significant differences in Raman bands intensity were found for sphingomyelin, acetyl CoA, methionine, DNA, RNA, phenylalanine, tryptophan, carotenoids, tyrosine, arginine, leucine, amide I and III, and phospholipids. The H NMR analysis showed reduced levels of D-glucose, L-proline, L-tyrosine, glycine, and anserine in HDP, while levels of 2-hydroxyvalerate, polyunsaturated fatty acids, and very-low-density lipoprotein (VLDL) were increased. H NMR results were able to assign an unknown sample to either the control or HDP groups at a precision of 88.3% using orthogonal partial least squares discriminant analysis and 87% using logistic regression analysis. Our results suggested that an analysis of exosome-enriched plasma could provide an initial assessment of placental function at the maternal-fetal interface and aid HDP diagnosis, prognosis, and treatment, as well as to detect novel, early biomarkers for HDP.
妊娠期高血压疾病与早产、死产以及孕产妇发病和死亡密切相关。妊娠期高血压疾病（HDP）通常在妊娠20周后才被发现，这限制了早期干预。胎盘通过释放细胞外囊泡（尤其是外泌体）等机制进行通讯，直接负责调节局部和全身生理功能。在本研究中，我们推测，对富含外泌体的母体血浆进行分析，可以为在妊娠早期更早诊断HDP提供一种更具针对性和适用性的方法。因此，在妊娠第20至24周期间收集了24名孕妇（11名对照组，13名HDP患者）的外周血血浆，并进行离心以富集外泌体。对富含外泌体的血浆样本进行拉曼光谱分析和质子核磁共振代谢组学（H NMR）分析。主成分分析（PCA）和正交偏最小二乘判别分析（OPLS-DA）用于分析600 - 1800 cm光谱区域的拉曼数据，以确定其区分不同组别的潜力。使用主成分分析，我们能够区分这两组，前三个主成分解释了所有方差的89%。在HDP患者中，鞘磷脂、乙酰辅酶A、蛋氨酸、DNA、RNA、苯丙氨酸、色氨酸、类胡萝卜素、酪氨酸、精氨酸、亮氨酸、酰胺I和III以及磷脂的拉曼谱带强度存在最显著差异。H NMR分析显示，HDP患者中D-葡萄糖、L-脯氨酸、L-酪氨酸、甘氨酸和鹅肌肽水平降低，而2-羟基戊酸、多不饱和脂肪酸和极低密度脂蛋白（VLDL）水平升高。使用正交偏最小二乘判别分析，H NMR结果能够以88.3%的精度将未知样本归为对照组或HDP组，使用逻辑回归分析的精度为87%。我们的结果表明，对富含外泌体的血浆进行分析可以对母胎界面的胎盘功能进行初步评估，并有助于HDP的诊断、预后和治疗，以及检测新的HDP早期生物标志物。
