In vitro model suggests oxidative stress involved in keratoconus disease.
In vitro model suggests oxidative stress involved in keratoconus disease.
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DOI：
10.1038/srep04608
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发表时间：
2014-04-09
影响因子：
4.6
通讯作者：
Zieske JD
中科院分区：
文献类型：
--
作者：
Karamichos D;Hutcheon AE;Rich CB;Trinkaus-Randall V;Asara JM;Zieske JD
圆锥角膜(KC)影响1：2000人，是一种角膜变薄并呈圆锥形的疾病。晚期KC需要手术来维持视力。氧化应激在KC中的作用尚不清楚。我们的目的是确定人角膜基质细胞(HCKs)、成纤维细胞(HCFs)和圆锥角膜细胞(HKCs)之间的氧化应激水平。细胞在2D和3D系统中培养。用维生素C(VitC)和转化生长因子β3(T3)刺激自组装细胞外基质4周。不使用T3作为对照。采用定量逆转录聚合酶链式反应和代谢组学方法对样本进行分析。QRT-PCR结果显示，HKCs的I型和V型胶原表达水平较低，角质形成蛋白的表达水平较低，表明HKCs向肌成纤维细胞表型分化。III型胶原是纤维化的标志，在HKCs中上调。在每个条件下，我们用LC-MS/MS检测到目标250个代谢产物中的150多个，其中几个与氧化应激有关。HKCs的乳酸水平、乳酸/苹果酸和乳酸/丙酮酸比值升高，而精氨酸和谷胱甘肽/氧化谷胱甘肽比值降低。在2D和3D中都发现了类似的图案。我们的数据显示，与角质形成细胞相比，成纤维细胞表现出更强的氧化应激。此外，HKC细胞的表达水平最高，提示它们可能具有肌成纤维细胞表型。
Keratoconus (KC) affects 1:2000 people and is a disorder where cornea thins and assumes a conical shape. Advanced KC requires surgery to maintain vision. The role of oxidative stress in KC remains unclear. We aimed to identify oxidative stress levels between human corneal keratocytes (HCKs), fibroblasts (HCFs) and keratoconus cells (HKCs). Cells were cultured in 2D and 3D systems. Vitamin C (VitC) and TGF-β3 (T3) were used for 4 weeks to stimulate self-assembled extracellular matrix (ECM). No T3 used as controls. Samples were analyzed using qRT-PCR and metabolomics. qRT-PCR data showed low levels of collagen I and V, as well as keratocan for HKCs, indicating differentiation to a myofibroblast phenotype. Collagen type III, a marker for fibrosis, was up regulated in HKCs. We robustly detected more than 150 metabolites of the targeted 250 by LC-MS/MS per condition and among those metabolites several were related to oxidative stress. Lactate levels, lactate/malate and lactate/pyruvate ratios were elevated in HKCs, while arginine and glutathione/oxidized glutathione ratio were reduced. Similar patterns found in both 2D and 3D. Our data shows that fibroblasts exhibit enhanced oxidative stress compared to keratocytes. Furthermore the HKC cells exhibit the greatest level suggesting they may have a myofibroblast phenotype.
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影响因子：
2.8
作者：
GONDHOWIARDJO, TD;VANHAERINGEN, NJ;KIJLSTRA, A
通讯作者：
KIJLSTRA, A
影响因子：
3.3
作者：
Karamichos, D.;Hutcheon, A. E. K.;Zieske, J. D.
通讯作者：
Zieske, J. D.
影响因子：
2.8
作者：
Karicherla P;Aras S;Aiyar A;Hobden JA
通讯作者：
Hobden JA
DOI：
10.1136/bjo.47.6.321
发表时间：
1963-06-01
期刊：
The British journal of ophthalmology
影响因子：
--
作者：
CULLEN, J F;BUTLER, H G
通讯作者：
BUTLER, H G
影响因子：
2.5
作者：
Dean, Olivia M.;van den Buuse, Maarten;Bush, Ashley I.
通讯作者：
Bush, Ashley I.