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The Chinese Journal of Dental Research

Year 2001
Volume 4 , Issue 4

Pages: 32 - 38

Induction of PGE2 Production and Cyclooxygenase-2 Gene Expression in Human Gingival Fibroblasts Stimulated with Lipopolysaccharide Isolated from Porphyromonas gingivalis and Prevotella Intermedia

Xiaozhen Jia/Mingwen Fan/Zhi Chen/Zhuan Bian/Chengzhang Li

Objective: To investigate the contribution of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) to PGE2 production in human gingival fibroblast cells stimulated with lipopolysaccharide from periodontopathogenic bacteria. Methods: Lipopolysaccharide was isolated from Porphyromonas gingivalis and Prevotella intermedia by the phenol-water method. Human gingival fibroblast cells were stimulated with lipopolysaccharide in different concentration for different time periods. The levels of PGE2 in the culture medium were measured by radioimmunoassay. The expression of COX-2 protein was observed by immunocytochemical staining; COX-1 and COX-2 mRNA expression were examined by semi-quantitative RT-PCR analysis. Results: The production of PGE2 in human gingival fibroblast cells was increased in a time- and dose-dependent manner (P < .01, P < .01). Immunocytochemical staining showed the positive expression of COX-2 protein in stimulated cells around the unstained cell nucleus, while no expression was found in unstimulated cells. The results by RT-PCR demonstrated COX-2 mRNA expression was increased after stimulation with P gingivalis and P intermedia-lipopolysaccharide. In contrast, COX-1 mRNA were not affected in either stimulated or control cells. Conclusion: P gingivalis and P intermedia-lipopolysaccharide can stimulate human gingival fibroblast cells to produce PGE2 via COX-2 gene expression in a dose- and time-dependent manner. The results indicate that COX-2 might be an important modulator in the process of periodontal destruction.



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