COLLAGEN-SYNTHESIS BY NORMAL AND FIBROTIC HUMAN-LUNG FIBROBLASTS AND THE EFFECT OF TRANSFORMING GROWTH FACTOR-BETA
COLLAGEN-SYNTHESIS BY NORMAL AND FIBROTIC HUMAN-LUNG FIBROBLASTS AND THE EFFECT OF TRANSFORMING GROWTH FACTOR-BETA
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DOI：
10.1164/ajrccm/140.1.95
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发表时间：
1989-07-01
期刊：
影响因子：
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通讯作者：
NARAYANAN, AS
中科院分区：
文献类型：
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作者：
RAGHU, G;MASTA, S;NARAYANAN, AS
胶原积聚是肺纤维化和其他纤维化病变的主要特征。我们研究了从正常和纤维化人肺样本培养的成纤维细胞中胶原的合成，并评估了它如何受到转化生长因子-β的影响。（TGF-β）。成纤维细胞获自正常和纤维化成人肺（n = 11;正常= 6，特发性肺纤维化= 5）。将它们暴露于TGF-β。用[3 H]脯氨酸和[3 H]甘氨酸脉冲标记。以细菌胶原酶敏感放射性来测量胶原蛋白的产生，并使用标记的前胶原α 1 [1] cDNA克隆HF 677作为探针通过溶液杂交测定来确定胶原蛋白mRNA水平。胶原蛋白的合成I，III，V型进行了评估后，通过DEAE-纤维素层析和SDS-聚丙烯酰胺凝胶电泳分离。结果表明，正常和纤维化肺成纤维细胞合成相似量的胶原。I型是由正常和纤维化细胞类型合成的主要胶原种类，并且I型、III型和V型胶原的相对比例在两种细胞类型中相似。TGF-.beta.导致胶原蛋白产生和胶原蛋白mRNA水平刺激增加2 - 4倍，并且在正常和纤维化肺成纤维细胞的反应中没有检测到差异。所有类型的胶原蛋白都被TGF-β刺激。TGF-.beta.不增加成纤维细胞增殖，并且大多数正常和纤维化肺细胞暴露于TGF-β。细胞周期仍处于G1期。我们的结论是，正常和纤维化的人肺成纤维细胞合成类似数量的胶原蛋白。TGF-.beta.在两种细胞株中刺激胶原蛋白产生的程度相同，并且在这两种成纤维细胞类型中，胶原蛋白产生的变化似乎是通过胶原蛋白mRNA水平介导的。
Collagen accumulation is a major feature of pulmonary fibrosis and other fibrotic lesions. We have studied the synthesis of collagens in fibroblasts cultured from normal and fibrotic human lung specimens and evaluated how it is affected by transforming growth factor-.beta. (TGF-.beta.). Fibroblasts were obtained from normal and fibrotic adult human lungs (n = 11; normal = 6, idiopathic pulmonary fibrosis = 5). They were exposed to TGF-.beta. and pulse-labeled with [3H]proline and [3H]glycine. Collagen production was measured as bacterial collagenase-susceptible radioactivity, and collagen mRNA levels were determined by a solution hybridization assay using labeled procollagen .alpha.1[1] cDNA clone HF677 as probe. Synthesis of collagen types I, III, and V were assessed after separating them by DEAE-cellulose chromatography and SDS-polyacrylamide gel electrophoresis. The results showed that both normal and fibrotic lung fibroblasts synthesized similar amounts of collagen. Type I was the major collagen species synthesized by both normal and fibrotic cell types, and the relative proportion of type I, III, and V collagens was similar in both cell types. TGF-.beta. caused a two to fourfold increase in stimulation of collagen production and collagen mRNA levels, and no differences were detected in the response of normal and fibrotic lung fibroblasts. All collagen types were stimulated by the TGF-.beta.. TGF-.beta. did not increase fibroblast proliferation and the majority of normal and fibrotic lung cells exposed to TGF-.beta. remained in G1 phase of the cell cycle. We conclude that fibroblasts of normal and fibrotic human lungs synthesize similar amounts of collagens. TGF-.beta. stimulates collagen production to the same extent in both cell strains and, in both of these fibroblast types, changes in collagen production appear to be mediated through collagen mRNA levels.