Human β-defensin-3 up-regulates cyclooxygenase-2 expression and prostaglandin E2 synthesis in human gingival fibroblasts
Chotjumlong P, Khongkhunthian S, Ongchai S, Reutrakul V, Krisanaprakornkit S. Human β-defensin-3 up-regulates cyclooxygenase-2 expression and prostaglandin E2synthesis in human gingival fibroblasts. J Periodont Res 2010; 45: 464-470. © 2010 John Wiley & Sons A/S Background and Objective: Oral ep...
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th-cmuir.6653943832-507372018-09-04T04:44:56Z Human β-defensin-3 up-regulates cyclooxygenase-2 expression and prostaglandin E2 synthesis in human gingival fibroblasts P. Chotjumlong S. Khongkhunthian S. Ongchai V. Reutrakul Suttichai Krisanaprakornkit Dentistry Chotjumlong P, Khongkhunthian S, Ongchai S, Reutrakul V, Krisanaprakornkit S. Human β-defensin-3 up-regulates cyclooxygenase-2 expression and prostaglandin E2synthesis in human gingival fibroblasts. J Periodont Res 2010; 45: 464-470. © 2010 John Wiley & Sons A/S Background and Objective: Oral epithelial cells express three antimicrobial peptide human β-defensins (hBDs) that have previously been demonstrated to exert proinflammatory effects on various immune cells. We wanted to examine whether hBDs could induce cyclooxygenase-2 (COX-2) expression and prostaglandin E2 (PGE2) synthesis in non-immune cells, such as human gingival fibroblasts. Material and Methods: Cultured fibroblasts were treated with different concentrations of hBD-1, -2, -3 or interleukin-1β, as a positive control, for various times, in the presence or absence of NS-398, a specific COX-2 inhibitor. The levels of COX-1 and COX-2 mRNA expression were analyzed using RT-PCR and real-time PCR. Whole cell lysates were analyzed for COX-1 and COX-2 protein expression by western blotting. Cell-free culture supernatants were assayed for PGE2 levels by ELISA. The lactate dehydrogenase assay was performed to determine the cytotoxicity of hBDs. Results: Ten and 40 μg/mL of hBD-3 up-regulated COX-2 mRNA and protein expression, consistent with COX-2 up-regulation by interleukin-1β, whereas hBD-1 and hBD-2 did not. However, COX-1 mRNA and protein were constitutively expressed. The time-course study revealed that hBD-3 up-regulated COX-2 mRNA and protein expression at 6 and 12 h, respectively. Consistent with COX-2 up-regulation, 10 and 40 μg/mL of hBD-3 significantly increased PGE2 levels in cell-free culture supernatants (p < 0.05), and this was inhibited by NS-398 in a dose-dependent manner. Neither of the hBD concentrations tested in this study was toxic to the cells. Conclusion: These findings indicate that epithelial human β-defensin-3 functions as a proinflammatory mediator in controlling arachidonic acid metabolism in underlying fibroblasts. © 2010 John Wiley & Sons A/S. 2018-09-04T04:44:56Z 2018-09-04T04:44:56Z 2010-08-01 Journal 16000765 00223484 2-s2.0-77954859999 10.1111/j.1600-0765.2009.01259.x https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=77954859999&origin=inward http://cmuir.cmu.ac.th/jspui/handle/6653943832/50737 |
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Dentistry P. Chotjumlong S. Khongkhunthian S. Ongchai V. Reutrakul Suttichai Krisanaprakornkit Human β-defensin-3 up-regulates cyclooxygenase-2 expression and prostaglandin E2 synthesis in human gingival fibroblasts |
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Chotjumlong P, Khongkhunthian S, Ongchai S, Reutrakul V, Krisanaprakornkit S. Human β-defensin-3 up-regulates cyclooxygenase-2 expression and prostaglandin E2synthesis in human gingival fibroblasts. J Periodont Res 2010; 45: 464-470. © 2010 John Wiley & Sons A/S Background and Objective: Oral epithelial cells express three antimicrobial peptide human β-defensins (hBDs) that have previously been demonstrated to exert proinflammatory effects on various immune cells. We wanted to examine whether hBDs could induce cyclooxygenase-2 (COX-2) expression and prostaglandin E2 (PGE2) synthesis in non-immune cells, such as human gingival fibroblasts. Material and Methods: Cultured fibroblasts were treated with different concentrations of hBD-1, -2, -3 or interleukin-1β, as a positive control, for various times, in the presence or absence of NS-398, a specific COX-2 inhibitor. The levels of COX-1 and COX-2 mRNA expression were analyzed using RT-PCR and real-time PCR. Whole cell lysates were analyzed for COX-1 and COX-2 protein expression by western blotting. Cell-free culture supernatants were assayed for PGE2 levels by ELISA. The lactate dehydrogenase assay was performed to determine the cytotoxicity of hBDs. Results: Ten and 40 μg/mL of hBD-3 up-regulated COX-2 mRNA and protein expression, consistent with COX-2 up-regulation by interleukin-1β, whereas hBD-1 and hBD-2 did not. However, COX-1 mRNA and protein were constitutively expressed. The time-course study revealed that hBD-3 up-regulated COX-2 mRNA and protein expression at 6 and 12 h, respectively. Consistent with COX-2 up-regulation, 10 and 40 μg/mL of hBD-3 significantly increased PGE2 levels in cell-free culture supernatants (p < 0.05), and this was inhibited by NS-398 in a dose-dependent manner. Neither of the hBD concentrations tested in this study was toxic to the cells. Conclusion: These findings indicate that epithelial human β-defensin-3 functions as a proinflammatory mediator in controlling arachidonic acid metabolism in underlying fibroblasts. © 2010 John Wiley & Sons A/S. |
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author |
P. Chotjumlong S. Khongkhunthian S. Ongchai V. Reutrakul Suttichai Krisanaprakornkit |
author_facet |
P. Chotjumlong S. Khongkhunthian S. Ongchai V. Reutrakul Suttichai Krisanaprakornkit |
author_sort |
P. Chotjumlong |
title |
Human β-defensin-3 up-regulates cyclooxygenase-2 expression and prostaglandin E2 synthesis in human gingival fibroblasts |
title_short |
Human β-defensin-3 up-regulates cyclooxygenase-2 expression and prostaglandin E2 synthesis in human gingival fibroblasts |
title_full |
Human β-defensin-3 up-regulates cyclooxygenase-2 expression and prostaglandin E2 synthesis in human gingival fibroblasts |
title_fullStr |
Human β-defensin-3 up-regulates cyclooxygenase-2 expression and prostaglandin E2 synthesis in human gingival fibroblasts |
title_full_unstemmed |
Human β-defensin-3 up-regulates cyclooxygenase-2 expression and prostaglandin E2 synthesis in human gingival fibroblasts |
title_sort |
human β-defensin-3 up-regulates cyclooxygenase-2 expression and prostaglandin e2 synthesis in human gingival fibroblasts |
publishDate |
2018 |
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https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=77954859999&origin=inward http://cmuir.cmu.ac.th/jspui/handle/6653943832/50737 |
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