Fibronectin synthesis in superficial and deep layers of normal articular cartilage.
Fibronectin synthesis in superficial and deep layers of normal articular cartilage.
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正常关节软骨浅层和深层的纤连蛋白合成。
DOI：
10.1002/art.1780390405
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发表时间：
1996
影响因子：
--
通讯作者：
Jasin,HE
中科院分区：
文献类型：
--
作者：
Hayashi,T;Abe,E;Jasin,HE
Objective.研究正常关节软骨浅层和深层纤维连接蛋白（FN）的分布和合成。采用牛和人关节软骨浅层和深层切片，提取FN，用放射免疫法测定其含量。软骨细胞也通过胶原酶消化分离用于FN提取和培养。浅层和深层软骨外植体培养和细胞因子的刺激。通过与35 S-甲硫氨酸孵育，对新合成的FN进行定量。在明胶琼脂糖柱上纯化FN，并通过聚丙烯酰胺凝胶电泳进一步表征。采用北方印迹法定量FN mRNA。新鲜分离的牛深层软骨细胞中FN含量是浅层软骨细胞的2.3 ± 0.2倍（P< 0.025）。深层软骨外植体含有1.2倍以上的FN比发现在浅表组织。深层软骨组织的FN合成量是浅层软骨组织的2.4倍（P< 0.01）。FN合成作为总蛋白合成的一部分，在深层外植体中显著高于浅层组织（P< 0.01）。分离培养的深层软骨细胞合成FN是表层细胞的1.89 ± 0.33倍（P< 0.05）。细胞因子刺激的浅表软骨外植体在FN合成方面没有反应。FNmRNA定量结果显示浅表和深部人群之间无显著性差异。由于FN起着重要的作用，在细胞粘附到受损的软骨表面，我们的研究结果表明，FN合成附近的关节软骨表面的调制可能是一个因素，阻碍血管翳入侵后，炎症损伤的关节。
Objective. To study the distribution and synthesis of fibronectin (FN) in superficial and deep layers of normal articular cartilage.Methods. Superficial and deep bovine and human articular cartilage slices were used to extract and quantitate FN by radioimmunoassay. Chondrocytes were also isolated by collagenase digestion for FN extraction and culture. Superficial and deep cartilage explants were cultured with and without stimulation by cytokines. Quantitation of newly synthesized FN was carried out by incubation with35S‐methionine. FN was purified on gelatin‐agarose columns and further characterized by polyacrylamide gel electrophoresis. FN messenger RNA (mRNA) was quantitated by Northern blot analysis.Results. Freshly isolated bovine chondrocytes from deep cartilage contained 2.3 ± 0.2 times more FN than was found in superficial cells (P< 0.025). Deep cartilage explants contained 1.2 times more FN than was found in superficial tissue. Explants obtained from deep cartilage synthesized 2.4 times more FN per cell than did superficial tissues (P< 0.01). FN synthesis as a fraction of total protein synthesis was significantly greater in deep explants (P< 0.01) compared with superficial tissues. Isolated deep chondrocytes in culture synthesized 1.89 ± 0.33‐fold more FN than did superficial cells (P< 0.05). Cytokine‐stimulated superficial cartilage explants failed to respond in terms of FN synthesis. FN mRNA quantitation showed no significant differences between superficial and deep populations.Conclusion. Since FN plays a major role in cell adhesion to damaged cartilage surfaces, our results suggest that modulation of FN synthesis near the articular surface of cartilage may be one of the factors that impede pannus invasion following an inflammatory insult to the joint.
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DOI：
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期刊：
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