EFFECT OF RELAXIN ON THE PHENOTYPE OF COLLAGENS SYNTHESIZED BY CULTURED RABBIT CHONDROCYTES
EFFECT OF RELAXIN ON THE PHENOTYPE OF COLLAGENS SYNTHESIZED BY CULTURED RABBIT CHONDROCYTES
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DOI：
10.1016/0167-4889(88)90119-x
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发表时间：
1988-11-18
期刊：
影响因子：
--
通讯作者：
CORVOL, MT
中科院分区：
文献类型：
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作者：
BONAVENTURE, J;DELATOUR, B;CORVOL, MT
通过测定总胶原产量和分析新合成胶原链的表型，研究了猪松弛素对原代培养的兔关节软骨细胞和生长板软骨细胞的影响。用2 μ g/ml松弛素处理单层关节和多层生长板软骨细胞24小时，对总DNA没有影响，也没有显著改变细胞分泌的[3 H]脯氨酸标记的胶原链的量。然而，聚丙烯酰胺凝胶电泳显示松弛素处理的软骨细胞相关的修改。观察到III型胶原的比例和对应于α 2 I链的条带的强度显著增加。CNBr-切割的分子的二维肽图谱表明被鉴定为α，一维凝胶上的II含有显著比例的α 1 I胶原蛋白链，如α 1 I溴化氰消化的肽的存在所证明的。松弛素处理后，这条带的强度增加。此外，通过狭缝印迹和北方印迹技术的总RNA分析显示，在与增加的松弛素浓度温育后，α 1 I和α 1 III mRNA水平的剂量依赖性刺激，但α 1 II mRNA的量没有变化。这些结果表明，当加入体外软骨细胞，松弛素调制的I型，II型和III型基因的表达，放大去分化过程。
The effect of porcine relaxin on rabbit articular and growth plate chondrocytes in primary culture was investigated by measurement of total collagen production and analysis of the phenotypes of newly synthesized collagen chains. A 24-h treatment of monolayer articular and multilayer growth plate chondrocytes with 2 .mu.g per ml relaxin had no effect on total DNA and did not significantly modify the amount of [3H]proline-labelled collagen chains secreted by the cells. However, polyacrylamide gel electrophoresis demonstrated relevant modifications in relaxin treated chondrocytes. A significant increase was observed in the proportion of type III collagen and in the intensity of the band corresponding to .alpha.2I chains. Two-dimensional peptide mapping of CNBr-cleaved molecules indicated that the band that was identified as .alpha., II on monodimensional gels contained a significant proportion of a1I collagen chains, as demonstrated by the presence of .alpha.1I cyanogen bromide-degested peptides. The intensity of this band was increased by relaxin treatment. Furthermore, total RNA analysis by slot blot and Northern blot techniques showed a dose-dependent stimulation of .alpha.1I and .alpha.1III mRNA levels after incubation with increased relaxin concentrations, but no change in the amount of .alpha.1II mRNA. These results suggested that when added to cartilage cells in vitro, relaxin modulated the expression of type I, type II and type III genes by amplifying the dedifferentiation process.