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中的一种，由成纤维细胞产生，对结缔组织的稳定性起重要作用。牙周病时，这些细胞基质可能受到细菌成分或细胞因子的刺激。为了了解这种变化的机制，我们研究了LPS和/或IL-1对培养成纤维细胞纤维连接蛋白和胶原蛋白产生的影响。成纤维细胞来源于2例健康牙龈和健康牙周韧带，1例炎症牙龈。这些成纤维细胞在含有10%胎牛血清的DMEM中培养。采用热-酚-水法分别从龈缝沉积物分离的牙龈假单胞菌381、中间假单胞菌ATCC 25611和牙龈假单胞菌中提取LPS。达到融合后，将成纤维细胞与重组IL-1和/或LPS分别孵育48小时。ELISA法检测培养基中纤维连接蛋白和胶原蛋白水平。1名受试者的健康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都有所增加，但另一名受试者的健康牙龈成纤维细胞水平在所有刺激下都有所下降。2例健康牙周韧带和炎症牙龈成纤维细胞的纤维连接蛋白和胶原蛋白水平均在所有刺激下升高。与LPS或IL-1相比，LPS和IL-1的刺激产生了更高水平的纤维连接蛋白或胶原蛋白。这些结果表明，来自不同对象的成纤维细胞在刺激作用下表现出不同的效果。

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.