Proteomic analysis of human osteoarthritic chondrocytes reveals protein changes in stress and glycolysis
Proteomic analysis of human osteoarthritic chondrocytes reveals protein changes in stress and glycolysis
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DOI：
10.1002/pmic.200700249
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发表时间：
2008-02-01
期刊：
影响因子：
3.4
通讯作者：
Blanco, Francisco J.
中科院分区：
文献类型：
--
作者：
Ruiz-Romero, Cristina;Carreira, Vanessa;Blanco, Francisco J.
骨关节炎（OA）以软骨退化为特征。软骨细胞是成熟软骨中唯一存在的细胞类型，它在软骨完整性的控制中起重要作用。本研究的目的是通过蛋白质组学方法分析OA软骨细胞的特征变化，并鉴定新的cia相关蛋白。从10例接受关节置换术的CIA患者和10例无关节疾病史的供体软骨中分离软骨细胞。全细胞蛋白用2-DE分离，并用SYPRO Ruby染色。用PDQuest 7.3.1软件分析OA和正常软骨细胞蛋白2-DE凝胶的蛋白表达模式。通过MALDI-TOF或MALDI-TOF/ TOF ms鉴定oa相关蛋白，Western blotting检测细胞中的ANXA1、GSTO1、GRP78和HSP90 β，免疫组化检测组织软骨中的HSP90 β。结果显示，2-DE凝胶中平均存在700个蛋白点。与正常软骨细胞相比，OA细胞有19个蛋白斑点显著增加（比值OA:N >= 2.0, p < 0.05）， CIA细胞有9个蛋白斑点减少（比值OA:N）
Osteoarthritis (OA) is characterized by cartilage degradation. The chondrocyte is the only cell type present in mature cartilage, and it is important in the control of cartilage integrity. The aim of this study was to analyze, by a proteomic approach, the changes that are characteristic of OA chondrocytes, and to identify new CIA-related proteins. Chondrocytes were isolated from the cartilage of ten CIA patients undergoing joint replacement and ten donors with no history of joint disease. Whole-cell proteins were resolved by 2-DE and stained with SYPRO Ruby. Protein expression patterns of 2-DE gels from OA and normal chondrocyte proteins were analyzed with PDQuest 7.3.1 software. OA-related proteins were identified by MALDI-TOF or MALDI-TOF/ TOF MS. The results were validated for ANXA1, GSTO1, GRP78, and HSP90 beta in cells by Western blotting and in tissue cartilage by immunohistochemistry Results showed an average of 700 protein spots that were present in the 2-DE gels. Compared to normal chondrocytes, 19 protein spots were found to be significantly increased in OA cells (ratio OA:N >= 2.0, p < 0.05), whereas nine were decreased in CIA chondrocytes (ratio OA:N