Department of Speech, Language, and Hearing Sciences, Purdue University, West Lafayette 47907, IN, United States.
Department of Comparative Pathobiology, Purdue University, West Lafayette 47907, IN, United States.
J Proteomics. 2022 Feb 10;252:104431. doi: 10.1016/j.jprot.2021.104431. Epub 2021 Nov 23.
Whole-body dehydration (i.e., systemic dehydration) leads to vocal fold tissue dehydration. Furosemide, a common diuretic prescribed to treat hypertension and edema-associated conditions, induces systemic dehydration. Furosemide also causes voice changes in human speakers, making this method of systemic dehydration particularly interesting for vocal fold dehydration studies. Our objective was to obtain a comprehensive proteome of vocal folds following furosemide-induced systemic dehydration. New Zealand White rabbits were used as the animal model and randomly assigned to euhydrated (control) or furosemide-dehydrated groups. Systemic dehydration, induced by injectable furosemide, was verified by an average body weight loss of -5.5% and significant percentage changes in blood analytes in the dehydrated rabbits compared to controls. Vocal fold specimens, including mucosa and muscle, were processed for proteomic analysis using label-free quantitation LC-MS/MS. Over 1600 proteins were successfully identified across all vocal fold samples; and associated with a variety of cellular components and ubiquitous cell functions. Protein levels were compared between groups showing 32 proteins differentially regulated (p ≤ 0.05) in the dehydrated vocal folds. These are mainly involved with mitochondrial translation and metabolism. The downregulation of proteins involved in mitochondrial metabolism in the vocal folds suggests a mechanism to prevent oxidative stress associated with systemic dehydration. SIGNIFICANCE: Voice disorders affect different population demographics worldwide with one in 13 adults in the United States reporting voice problems annually. Vocal fold systemic hydration is clinically recognized for preventing and treating voice problems and depends on optimal body hydration primarily achieved by water intake. Herein, we use the rabbit as a translatable animal model, and furosemide as a translatable method of systemic dehydration, to reveal a comprehensive proteomic profile of vocal fold mucosa and muscle in response to systemic dehydration. The significant subset of proteins differentially regulated due to furosemide-induced dehydration offer novel insights into the molecular mechanisms of systemic dehydration in the vocal folds. These findings also deepen our understanding of changes to tissue biology after diuretic administration.
全身性脱水（即全身脱水）可导致声带组织脱水。呋塞米是一种常用于治疗高血压和与水肿相关疾病的利尿剂，可引起全身性脱水。呋塞米还会导致人类说话者的声音发生变化，因此这种全身性脱水方法对于声带脱水研究特别有趣。我们的目的是获得呋塞米诱导的全身性脱水后声带的全面蛋白质组图谱。新西兰白兔被用作动物模型，并随机分为正常水合（对照）或呋塞米脱水组。通过注射呋塞米诱导的全身性脱水，通过脱水兔与对照相比平均体重减轻 5.5%和血液分析物的显著百分比变化来验证。对包括粘膜和肌肉在内的声带标本进行蛋白质组分析，使用无标签定量 LC-MS/MS。在所有声带样本中成功鉴定了超过 1600 种蛋白质；并与各种细胞成分和普遍存在的细胞功能相关。对两组之间的蛋白质水平进行比较，显示 32 种蛋白质在脱水的声带中差异调节（p≤0.05）。这些主要涉及线粒体翻译和代谢。在声带中与线粒体代谢有关的蛋白质下调表明了一种预防与全身性脱水相关的氧化应激的机制。意义：世界各地的不同人群都受到声音障碍的影响，美国每 13 名成年人中就有 1 人每年报告有声音问题。声带全身水合作用被临床认可为预防和治疗声音问题，主要取决于通过饮水达到最佳的全身水合作用。在此，我们使用兔作为可翻译的动物模型，并用呋塞米作为可翻译的全身性脱水方法，揭示了声带粘膜和肌肉对全身性脱水的全面蛋白质组图谱。由于呋塞米诱导的脱水而差异调节的显著蛋白质子集为声带全身脱水的分子机制提供了新的见解。这些发现还加深了我们对利尿剂给药后组织生物学变化的理解。
