Effects of LPS and/or IL-1 on fibronectin and collagen synthesis in cultured fibroblasts

纤维连接蛋白和胶原是细胞基质之一，由成纤维细胞产生，对结缔组织的稳定性起重要作用。这些细胞基质可能受到牙周病中细菌或细胞因子成分的刺激。为了了解这种变化的机制，我们研究了LPS和/或IL-1对培养的成纤维细胞产生纤维连接蛋白和胶原的影响。成纤维细胞从2名受试者的健康牙龈和健康牙周膜以及1名患者的发炎牙龈中获得。将这些成纤维细胞在含有10%胎牛血清的DMEM中培养。采用热酚水法从牙龈卟啉单胞菌381、中间卟啉单胞菌ATCC 25611和从龈沟沉积物中分离的牙龈卟啉单胞菌中提取LPS。在达到汇合后，将成纤维细胞在存在或不存在重组IL-1和/或LPS的情况下进一步孵育48小时。用ELISA法测定培养液中纤维连接蛋白和胶原的含量。来自1名受试者的健康牙龈成纤维细胞的纤连蛋白和胶原蛋白的水平在所有刺激的存在下增加，但另一名受试者的健康牙龈成纤维细胞的水平通过所有刺激降低。2例健康牙周膜和炎症牙龈成纤维细胞的纤维连接蛋白和胶原蛋白水平在所有刺激下均升高。LPS和IL-1的刺激产生更高水平的纤连蛋白或胶原蛋白相比，LPS或IL-1单独。这些结果表明，不同受试者的成纤维细胞在刺激后表现出不同的效应。

fibronectin and collagen, one of the cell matrix, is produced by fibroblasts and plays an important role for stability of connective tissues. These cell matrix may be stimulated with components of bacteria or cytokines in periodontal disease. To know the mechanism of this change, we studied effects of LPS and/or IL-1 on production of fibronectin and collagen by cultured fibroblasts. Fibroblasts were obtained from healthy gingiva and healthy periodontal ligament of 2 subjects, and from inflamed gingiva of 1 patient. These fibroblasts were cultured in DMEM containing 10% Fetal Bovine Serum. LPS were extracted from P. gingivalis 381, P. intermedia ATCC 25611 and P. gingivalis isolated from gingival crevice deposits by the hot-phenol-water method. After reaching confluent, the fibroblasts were further incubated with or without present of recombinant IL-1 and/or the LPS for 48 hours. Levels of fibronectin and collagen in culture medium were measured by ELISA. The levels of fibronectin and collagen of healthy gingival fibroblasts from 1 subject were increased in presence of all stimulation, but the levels of healthy gingival fibroblasts of another subject were decreased by all stimulation. The levels of fibronectin and collagen of healthy periodontal ligament from 2 subjects and inflamed gingival fibroblasts were increased by all stimulation. The stimulation of LPS and IL-1 produced higher levels of fibronectin or collagen as compared to those of LPS or IL-1 only. These findings were suggested that fibroblasts from difference subjects showed difference effects by stimulation.