Title: | Oral malodorous compound triggers mitochondrial-dependent apoptosis and causes genomic DNA damage in human gingival epithelial cells |
Author(s): | Calenic B; Yaegaki K; Murata T; Imai T; Aoyama I; Sato T; Ii H; |
Address: | "Department of Oral Health, Nippon Dental University, Tokyo, Japan" |
DOI: | 10.1111/j.1600-0765.2008.01199.x |
ISSN/ISBN: | 1600-0765 (Electronic) 0022-3484 (Linking) |
Abstract: | "BACKGROUND AND OBJECTIVE: Volatile sulfur compounds are the main compounds causing halitosis. One of these compounds, hydrogen sulfide (H(2)S), which is responsible for physiological halitosis, is reported also to have periodontal pathogenic activities. Hydrogen sulfide has been shown to activate the apoptotic process in different tissues. Apoptosis plays an important role in the development of periodontitis. The aim of this study was to determine whether H(2)S causes apoptosis in human gingival epithelial cells and to examine the cellular signaling pathway initiating the process. MATERIAL AND METHODS: Human gingival epithelial cells were incubated with 50 ng/mL H(2)S in air contining 5% CO(2) for 24, 48 or 72 h. To detect apoptosis, the cells were stained with annexin V and 7-amino actinomycin D, and analyzed using flow cytometry. Reactive oxygen species, mitochondrial membrane depolarization and release of cytochrome C into the cytosol were assessed using flow cytometry and enzyme-linked immunosorbent assay. Activity levels for the key apoptotic enzymes caspase-9, -8 and -3 were also determined. Genomic DNA damage was detected using single-cell gel electrophoresis. RESULTS: Apoptosis was significantly increased to 24.5 +/- 5.7 at 24 h and 41.5 +/- 8.9% at 48 h (p < 0.01). Reactive oxygen species were enhanced and mitochondrial membrane depolarization was collapsed. Cytochrome C release was dramatically increased (0.12 +/- 0.02 vs. 0.02 +/- 0.01 at 24 h and 0.21 +/- 0.02 vs. 0.02 +/- 0.01 ng/mL at 48 h; p < 0.05). Caspase-9 and -3 were strongly activated, while caspase-8 activity remained low. The percentage of DNA strand breaks increased, especially at 48 h. CONCLUSION: Hydrogen sulfide induces apoptosis in human gingival epithelial cells by activating the mitochondrial pathway" |
Keywords: | "Annexin A5 Apoptosis/*drug effects Caspase 3/analysis Caspase 8/analysis Caspase 9/analysis Cell Line Cells, Cultured Comet Assay Cytochromes c/analysis *DNA Damage Dactinomycin/analogs & derivatives Epithelial Cells/drug effects/ultrastructure Flow Cytom;" |
Notes: | "MedlineCalenic, B Yaegaki, K Murata, T Imai, T Aoyama, I Sato, T Ii, H eng Research Support, Non-U.S. Gov't 2009/07/16 J Periodontal Res. 2010 Feb; 45(1):31-7. doi: 10.1111/j.1600-0765.2008.01199.x. Epub 2009 Jul 8" |