Vuorio T, Mäkelä J K, Vuorio E
J Cell Biochem. 1985;28(2):105-13. doi: 10.1002/jcb.240280204.
Fibroblasts cultured from affected skin areas of five patients with cutaneous scleroderma were found to produce increased amounts of collagen when compared with nonaffected control cells. Total RNA was isolated from the cultures and analyzed for its level of pro alpha 1 (I)collagen mRNA by hybridization of RNA blots with a cloned cDNA probe. The levels of pro alpha 1 (I)collagen mRNAs relative to total RNA were two- to sixfold higher in the samples from affected cells, accounting for the increased synthesis of type I collagen. Cytoplasmic dot hybridizations were performed to measure the cellular content of pro alpha 1 (I)collagen mRNA: up to ninefold increases in the level of this mRNA per cell were found. Upon subculturing, scleroderma fibroblasts were found to reduce gradually the increased synthesis of collagen to the level of nonaffected controls by the tenth passage. The levels of type I collagen mRNAs were also reduced, but more slowly. The results suggest that in scleroderma fibroblasts the genes for type I collagen are activated at procollagen mRNA level or that they are more stable and that the activating factors are lost during prolonged cell culture because cells from affected areas lose their activated state.
与未受影响的对照细胞相比，从五名皮肤硬皮病患者受影响皮肤区域培养的成纤维细胞产生的胶原蛋白量增加。从培养物中分离出总RNA，并通过用克隆的cDNA探针杂交RNA印迹分析其原α1（I）型胶原蛋白mRNA的水平。与总RNA相比，受影响细胞样本中原α1（I）型胶原蛋白mRNA的水平高两到六倍，这解释了I型胶原蛋白合成的增加。进行细胞质斑点杂交以测量原α1（I）型胶原蛋白mRNA的细胞含量：发现每个细胞中该mRNA的水平最多增加九倍。传代培养后，发现硬皮病成纤维细胞在第十代时逐渐将增加的胶原蛋白合成降低到未受影响对照的水平。I型胶原蛋白mRNA的水平也降低了，但更缓慢。结果表明，在硬皮病成纤维细胞中，I型胶原蛋白基因在原胶原蛋白mRNA水平被激活，或者它们更稳定，并且在长时间的细胞培养过程中激活因子丢失，因为来自受影响区域的细胞失去了它们的激活状态。
