The overexpression of cyclo-oxygenase-2 in chronic periodontitis

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The overexpression of cyclo-oxygenase-2 in chronic periodontitis FAN ZHANG, M.D., Ph.D., STEVEN P. ENGEBRETSON, D.M.D., M.S., RAKHI S. MORTON, D.D.S., M.S., PAUL F. CAVANAUGH, Ph.D., KOTHA SUBBARAMAIAH, Ph.D., ANDREW J. DANNENBERG, M.D. The Journal of the American Dental Association Volume 134, Issue 7, Pages 861-867 (July 2003) DOI: 10.14219/jada.archive.2003.0284 Copyright © 2003 American Dental Association Terms and Conditions

Figure 1 Levels of cyclo-oxygenase-2, or COX-2, mRNA are increased in gingival tissue from patients with chronic periodontitis, or CP. A: A representative quantitative reverse transcriptase–polymerase chain reaction, or RT-PCR, in a case of CP. Total RNA (0.5 micrograms) and known concentrations (0.001–0.05 pg) of hCOX-2 mimic internal control are competing for a fixed amount of hCOX-2 primer in each reaction, giving rise to relative proportions of 569-bp internal control product and 724-bp target COX-2 cDNA product. B: Quantitative RT-PCR was used to determine the amounts of COX-2 mRNA in eight samples of gingivae from healthy subjects (mean ± standard error of the mean, or SEM, 28.0 ± 9 .4 fg/μg of total RNA) and in 10 samples of gingivae from patients with CP (mean ± SEM, 99.3 ± 17.2 fg/μg of total RNA). The asterisk indicatesP< .01. The Journal of the American Dental Association 2003 134, 861-867DOI: (10.14219/jada.archive.2003.0284) Copyright © 2003 American Dental Association Terms and Conditions

Figure 2 Increased levels of cyclo-oxygenase-2, or COX-2, protein in gingival tissue from patients with chronic periodontitis, or CP. The authors performed immunoblotting on gingivae from three healthy volunteers and from four patients with CP. Tissue lysate protein (100 micrograms/lane) was loaded onto a 10 percent sodium dodecyl sulfate–polyacrylamide gel and electrophoresed. The immunoblot was probed sequentially with antibodies to COX-2 and β-actin. Ovine COX-2 was used as a standard. The Journal of the American Dental Association 2003 134, 861-867DOI: (10.14219/jada.archive.2003.0284) Copyright © 2003 American Dental Association Terms and Conditions

Figure 3 Effects of tumor necrosis factor alpha, or TNF-α, interleukin 1 beta, or IL-1β, and lipopolysaccharide, or LPS, on the expression of cyclo-oxygenase-2, or COX-2. Lysate protein was from RAW 264.7 (panel A) or 1483 (panel B) cells treated with vehicle (control), TNF-α, IL-1β or LPS for six hours. Cellular lysate protein (30 micrograms/lane) was loaded onto a 10 percent sodium dodecyl sulfate–polyacrylamide gel, electrophoresed and transferred onto nitrocellulose. Immunoblots were probed with antibodies specific for COX-2 and β-actin. ng/mL: Nanograms per milliliter. The Journal of the American Dental Association 2003 134, 861-867DOI: (10.14219/jada.archive.2003.0284) Copyright © 2003 American Dental Association Terms and Conditions

Figure 4 Schematic depicting the stimulation of cyclo-oxygenase-2, or COX-2, expression in the periodontium by paracrine and autocrine factors produced as part of the inflammatory cascade initiated by periodontal pathogens. IL-1β: Interleukin 1 beta. PGE2: Prostaglandin E2. TNF-α: Tumor necrosis factor alpha. The Journal of the American Dental Association 2003 134, 861-867DOI: (10.14219/jada.archive.2003.0284) Copyright © 2003 American Dental Association Terms and Conditions