Valentim-Coelho Cristina, Vaz Fátima, Antunes Marília, Neves Sofia, Martins Inês L, Osório Hugo, Feliciano Amélia, Pinto Paula, Bárbara Cristina, Penque Deborah
Laboratório de Proteómica, Departamento de Genética Humana, Instituto Nacional de Saúde Dr. Ricardo Jorge, 1649-016 Lisboa, Portugal.
ToxOmics-Centre of Toxicogenomics and Human Health, Universidade Nova de Lisboa, 1150-082 Lisboa, Portugal.
Antioxidants (Basel). 2020 Nov 26;9(12):1184. doi: 10.3390/antiox9121184.
In this study, we examined the effect of six months of positive airway pressure (PAP) therapy on Obstructive Sleep Apnea (OSA) red blood cell (RBC) proteome by two dimensional difference gel electrophoresis (2D-DIGE) - based proteomics followed by Western blotting (WB) validation. The discovered dysregulated proteins/proteoforms are associated with cell death, HO catabolic/metabolic process, stress response, and protein oligomerization. Validation by nonreducing WB was performed for peroxiredoxin-2 (PRDX2) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) by using antibodies against the sulfinylated/sulfonylated cysteine of these proteins to better evaluate their redox-oligomeric states under OSA and/or in response to PAP therapy. The results indicated that the redox-oligomeric state of GAPDH and PRDX2 involving overoxidation by sulfinic/sulfonic acids were differentially modulated in OSA RBC, which might be compromising RBC homeostasis. PAP therapy by restoring this modulation induced a higher oligomerization of overoxidized GAPDH and PRDX2 in some patients that could be associated with eryptosis and the chaperone "gain" of function, respectively. This varied response following PAP may result from the complex interplay between OSA and OSA metabolic comorbidity. Hence, information on the redox status of PRDX2 and GAPDH in RBC will help to better recognize OSA subtypes and predict the therapeutic response in these patients. GAPDH monomer combined with body mass index (BMI) and PRDX2 S-S dimer combined with homeostatic model assessment for insulin resistance (HOMA-IR) showed to be very promising biomarkers to predict OSA and OSA severity, respectively.
在本研究中，我们通过基于二维差异凝胶电泳（2D-DIGE）的蛋白质组学方法，随后进行蛋白质印迹（WB）验证，研究了为期六个月的气道正压通气（PAP）治疗对阻塞性睡眠呼吸暂停（OSA）红细胞（RBC）蛋白质组的影响。发现失调的蛋白质/蛋白质变体与细胞死亡、血红素加氧酶（HO）分解代谢/代谢过程、应激反应和蛋白质寡聚化有关。通过使用针对这些蛋白质的亚磺酰化/磺酰化半胱氨酸的抗体，对过氧化物酶2（PRDX2）和甘油醛-3-磷酸脱氢酶（GAPDH）进行非还原WB验证，以更好地评估它们在OSA情况下和/或对PAP治疗反应时的氧化还原寡聚状态。结果表明，在OSA红细胞中，GAPDH和PRDX2的氧化还原寡聚状态因亚磺酸/磺酸过度氧化而受到不同调节，这可能会损害红细胞的内环境稳定。PAP治疗通过恢复这种调节，在一些患者中诱导了过度氧化的GAPDH和PRDX2更高程度的寡聚化，这可能分别与红细胞凋亡和伴侣蛋白功能“获得”有关。PAP治疗后这种不同的反应可能是由于OSA与OSA代谢合并症之间复杂的相互作用所致。因此，关于红细胞中PRDX2和GAPDH氧化还原状态的信息将有助于更好地识别OSA亚型并预测这些患者的治疗反应。GAPDH单体与体重指数（BMI）相结合，PRDX2 S-S二聚体与胰岛素抵抗稳态模型评估（HOMA-IR）相结合，分别显示出是预测OSA和OSA严重程度非常有前景的生物标志物。
