Laboratorio de Hematobiología, Escuela Nacional de Medicina y Homeopatía, Instituto Politécnico Nacional, Mexico City, Mexico.
CONACyT Fellow-ICAT, Universidad Nacional Autónoma de México, Mexico City, Mexico.
Biosens Bioelectron. 2020 Jun 1;157:112151. doi: 10.1016/j.bios.2020.112151. Epub 2020 Mar 14.
Arterial hypertension (HTN) is a world health concern presenting difficulties for its early detection. It leads to cardiovascular and kidney complications that increase morbidity in adults. Overexpression in the epithelial sodium channel (ENaC) in membrane platelets can be related with the presence of HTN and thus can be used as a biomarker to detect this medical condition. Here, we propose a method for HTN diagnosis based on gold nanoparticles (GNPs) conjugated to an antibody against the ENaC present on platelets. These functionalized GNPs were analyzed by Zeta potential, dynamic light scattering, electron microscopy, and other spectroscopic techniques. To verify that the GNPs and α-ENaC antibodies formed conjugates (GNPs-antiENaC) that maintained their specificity to the target, we carried out an indirect immunofluorescence detection assay of GNPs-antiENaC bound to a secondary antibody labeled with a fluorophore. Our results show that the presence of GNPs increase the fluorescence intensity in platelets treated with GNPs-antiENaC conjugates. It is also observed a clear tendency of the fluorescence signal in platelets treated with the conjugates that could be used for discrimination between normotensive and hypertensive samples. The proposed assay can be implemented as a very sensitive routine test to diagnose HTN.
动脉高血压（HTN）是一个全球性的健康问题，其早期检测存在困难。它会导致心血管和肾脏并发症，使成年人的发病率增加。在膜血小板中上皮钠离子通道（ENaC）的过度表达可能与 HTN 的存在有关，因此可以作为检测这种疾病的生物标志物。在这里，我们提出了一种基于与血小板上存在的 ENaC 抗体结合的金纳米颗粒（GNPs）的 HTN 诊断方法。这些功能化的 GNPs 通过 Zeta 电位、动态光散射、电子显微镜和其他光谱技术进行分析。为了验证 GNPs 和 α-ENaC 抗体形成了保持其对靶特异性的缀合物（GNPs-antiENaC），我们进行了间接免疫荧光检测，检测与荧光标记的二级抗体结合的 GNPs-antiENaC。我们的结果表明，GNPs 的存在增加了用 GNPs-antiENaC 缀合物处理的血小板中的荧光强度。还观察到在用缀合物处理的血小板中荧光信号的明显趋势，可用于区分正常血压和高血压样本。所提出的测定法可作为诊断 HTN 的非常敏感的常规试验来实施。
