Biochemical comparison of fibroblast populations from different periodontal tissues: characterization of matrix protein and collagenolytic enzyme synthesis.
Biochemical comparison of fibroblast populations from different periodontal tissues: characterization of matrix protein and collagenolytic enzyme synthesis.
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不同牙周组织成纤维细胞群的生化比较：基质蛋白和胶原分解酶合成的表征。
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发表时间：
1988
期刊：
影响因子：
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通讯作者：
J. Sodek
中科院分区：
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作者：
K. Otsuka;S. Pitaru;C. Overall;J. Aubin;J. Sodek
为了比较不同牙周组织成纤维细胞对细胞外基质成分的表达，本实验从动物个体中分离培养大鼠臼齿牙周韧带成纤维细胞（RPL）和大鼠牙龈成纤维细胞（RGF）。以[35S]蛋氨酸为前体的脉冲追踪实验显示，两种细胞类型的早期传代细胞的融合群体合成了相似数量的胶原蛋白、纤维连接蛋白和SPARC/骨连接蛋白。在III型胶原蛋白的相对比例、前胶原蛋白的加工速度以及通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳观察到的少量未识别蛋白的合成方面，定性和定量的差异是明显的。两种细胞类型分泌的放射性标记蛋白中胶原蛋白占24-26%，I型胶原占主导地位，III型胶原（3-8% RGF, 8-18% RPL）和V型胶原含量较低（约1%）。前胶原在RPL细胞培养基中的加工速度比RGF细胞快，但在RPL和RGF的多层培养中均有所增加。在多层培养中，胶原TCA片段，指示组织胶原酶活性，也被确定。在这些培养中发现了活性和潜伏的组织胶原酶以及一种新的胶原溶解酶（基质金属内源性蛋白酶v）的潜伏形式，这种酶可以切割天然的TCA片段。在培养基中加入刀豆蛋白A （10(-6) M）或视黄酸（10(-5)M）刺激潜在的胶原溶解酶的分泌。RGF和RPL细胞均可合成胶原酶抑制剂。SPARC/骨连接蛋白是一种40千道尔顿的糖蛋白，占两种细胞分泌的放射性标记蛋白的0.5-1.0%。
To compare the expression of extracellular matrix components by fibroblasts from different periodontal tissues, rat molar periodontal ligament fibroblasts (RPL) and rat gingival fibroblasts (RGF) were isolated and cultured from individual animals. Pulse-chase experiments using [35S]methionine as a precursor revealed that confluent populations of early passage cells of both cell types synthesized similar amounts of collagen, fibronectin, and SPARC/osteonectin. Qualitative and quantitative differences were apparent in the relative proportions of type III collagen, in the rates of procollagen processing, and in the synthesis of a small number of unidentified proteins observed by sodium dodecyl sulphate--polyacrylamide gel electrophoresis. Collagen constituted 24-26% of the radiolabelled proteins secreted by both cell types, type I being the predominant collagen, with lower amounts of type III (3-8% RGF, 8-18% RPL) and type V (approximately 1%) collagens. Procollagen processing in the culture medium of RPL cells was more rapid than for RGF cells, but was increased in multilayered cultures of both RPL and RGF. In multilayered cultures, collagen TCA fragments, indicative of tissue collagenase activity, were also identified. Active and latent tissue collagenases and a latent form of a novel collagenolytic enzyme (matrix metalloendoproteinase-V) that cleaves native TCA fragments were demonstrated in these cultures. Addition of either concanavalin A (10(-6) M) or retinoic acid (10(-5) M) to the culture medium stimulated the secretion of the latent collagenolytic enzymes. Collagenase inhibitor was also synthesized by both RGF and RPL cells. SPARC/osteonectin, a 40-kilodalton glycoprotein, represented 0.5-1.0% of the secreted radiolabelled proteins of both cell types.