Suarez Maria Fernanda, Piqueras Maria Carmen, Correa Leandro, Esposito Evangelina, Barros Maria Fernanda, Bhattacharya Sanjoy K, Urrets-Zavalia Julio A, Serra Horacio M
CIBICI-CONICET, Faculty of Chemical Sciences, Department of Clinical Biochemistry, Universidad Nacional de Cordoba, Cordoba, Argentina.
Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida.
J Cell Biochem. 2017 Nov;118(11):3920-3931. doi: 10.1002/jcb.26045. Epub 2017 May 23.
Climatic droplet keratopathy (CDK) is an acquired degenerative disease predominantly affecting males over 40 years old. It results in progressive corneal opacities usually affecting both eyes. CDK is multifactorial and its etiology remains unknown. Our recent findings are consistent with CDK pathology being driven by environmental factors with oxidative stress playing an important role (e.g.,, contributing to lipid peroxidation) rather than climate factors. The changes in corneal lipid composition affected by environmental factors remain understudied. The purpose of this study was to systematically investigate phospholipids profile (phosphatidylcholine [PC] and phosphatidylserine [PS]) in corneas from CDK patients using tandem mass spectrometry. Samples from CDK areas and from non-affected areas were obtained from patients diagnosed with CDK who underwent cataract surgery, were subjected to lipid extraction using a modified Bligh and Dyer method; protein concentrations were determined using the Bradford's method. Lipids were identified and subjected to ratiometric quantification using TSQ Quantum Access Max triple quadrupole mass spectrometer, using appropriate class specific lipid standards. All phospholipid classes showed lower total amounts in affected areas compared to control areas from CDK's corneas. Comparative profiles of two phospholipid classes (PC, PS) between CDK areas and control areas showed several common species between them. We also found a few unique lipids that were absent in CDK areas compared to controls and vice versa. Lower amount of phospholipids in CDK areas compared to control areas could be attributed to the lipid peroxidation in the affected corneal regions as a consequence of increased oxidative stress. J. Cell. Biochem. 118: 3920-3931, 2017. © 2017 Wiley Periodicals, Inc.
气候性点状角膜病变（CDK）是一种获得性退行性疾病，主要影响40岁以上男性。它会导致渐进性角膜混浊，通常累及双眼。CDK是多因素的，其病因尚不清楚。我们最近的研究结果表明，CDK的病理学是由环境因素驱动的，氧化应激起着重要作用（例如，导致脂质过氧化），而不是气候因素。环境因素对角膜脂质成分的影响仍未得到充分研究。本研究的目的是使用串联质谱法系统地研究CDK患者角膜中的磷脂谱（磷脂酰胆碱[PC]和磷脂酰丝氨酸[PS]）。从接受白内障手术的CDK患者中获取CDK区域和未受影响区域的样本，采用改良的布利和戴尔方法进行脂质提取；使用布拉德福德法测定蛋白质浓度。使用适当的类别特异性脂质标准品，通过TSQ Quantum Access Max三重四极杆质谱仪鉴定脂质并进行比例定量。与CDK角膜的对照区域相比，所有磷脂类在受影响区域的总量均较低。CDK区域和对照区域之间两种磷脂类（PC、PS）的比较图谱显示它们之间有几种常见的种类。我们还发现了一些与对照相比在CDK区域不存在的独特脂质，反之亦然。与对照区域相比，CDK区域中磷脂含量较低可能归因于受影响角膜区域由于氧化应激增加而导致的脂质过氧化。《细胞生物化学杂志》118: 3920 - 3931, 2017。© 2017威利期刊公司。
