Age-related changes in cartilage endogenous osteogenic protein-1 (OP-1)
Age-related changes in cartilage endogenous osteogenic protein-1 (OP-1)
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DOI：
10.1016/s0925-4439(02)00158-8
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发表时间：
2002-11-20
影响因子：
6.2
通讯作者：
Kuettner, KE
中科院分区：
文献类型：
--
作者：
Chubinskaya, S;Kumar, B;Kuettner, KE
关节软骨修复能力差。该特征随着年龄增长和退行性关节疾病而加剧，导致活动度丧失和生活质量受损。本研究的重点是成骨蛋白-1（OP-1）及其作为关节软骨修复刺激因子的能力。本研究的目的是建立一种定量分析人关节软骨提取物中OP-1蛋白含量的方法，并研究正常成人关节软骨中OP-1 mRNA表达和蛋白水平随衰老的变化。从死亡后24小时内没有关节疾病史的供体的股骨髁解剖全层软骨。通过半定量逆转录-聚合酶链反应（RT-PCR）方法测定OP-1 mRNA的表达水平;通过新的夹心酶联免疫吸附试验（ELISA）检测OP-1蛋白的浓度;用各种抗OP-1抗体通过Western印迹法评价OP-1形式的定性变化。ELISA方法的灵敏度允许在软骨提取物中检测皮克量的OP-1。我们发现：（1）正常软骨中OP-1的浓度在体外OP-1的生物活性范围内;（2）在人成年关节软骨的老化过程中，OP-1蛋白和信使的水平显著降低（超过4倍; P < 0.05）。
Articular cartilage has a poor reparative capacity. This feature is exacerbated with aging and during degenerative joint conditions, contributing to loss of motion and impairment of quality of life. This study focused on osteogenic protein-1 (OP-1) and its ability to serve as a repair-stimulating factor in articular cartilage. The purpose of this work was to develop a quantitative method for the assessment of the content of OP-1 protein in extracts from human articular cartilage and to investigate the changes in OP-1 mRNA expression and protein levels with aging of normal adult cartilage. Full thickness cartilage was dissected from femoral condyles of donors with no history of joint disease within 24 h of death. Levels of OP-1 mRNA expression were measured by a semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) method; concentration of OP-1 protein was detected by new sandwich enzyme-linked immunosorbent assay (ELISA); qualitative changes in OP-1 forms were evaluated by Western blots with various anti-OP-1 antibodies. The sensitivity of the ELISA method allowed the detection of picogram quantities of OP-1 in cartilage extracts. We found that (1) concentration of OP-1 in normal cartilage is within the range of biological activity of OP-1 in vitro; and (2) during aging of human adult, articular cartilage, levels of OP-1 protein and message are dramatically reduced (more than 4-fold; p