Butyrate induces reactive oxygen species production and affects cell cycle progression in human gingival fibroblasts

M. C. Chang, Y. L. Tsai, Y. W. Chen, C. P. Chan, C. F. Huang, W. C. Lan, C. C. Lin, W. H. Lan, J. H. Jeng

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Background and Objective: Short-chain fatty acids, such as butyric acid and propionic acid, are metabolic by-products generated by periodontal microflora such as Porphyromonas gingivalis, and contribute to the pathogenesis of periodontitis. However, the effects of butyrate on the biological activities of gingival fibroblasts (GFs) are not well elucidated. Material and Methods: Human GFs were exposed to various concentrations of butyrate (0.5-16mm) for 24h. Viable cells that excluded trypan blue were counted. Cell cycle distribution of GFs was analyzed by propidium iodide-staining flow cytometry. Cellular reactive oxygen species (ROS) production was measured by flow cytometry using 2',7'-dichlorofluorescein (DCF). Total RNA and protein lysates were isolated and subjected to RT-PCR using specific primers or to western blotting using specific antibodies, respectively. Results: Butyrate inhibited the growth of GFs, as indicated by a decrease in the number of viable cells. This event was associated with an induction of G0/G1 and G2/M cell cycle arrest by butyrate (4-16mm) in GFs. However, no marked apoptosis of GFs was noted in this experimental condition. Butyrate (>2mm) inhibited the expression of cdc2, cdc25C and cyclinB1 mRNAs and reduced the levels of Cdc2, Cdc25C and cyclinB1 proteins in GFs, as determined using RT-PCR and western blotting, respectively. This toxic effect of butyrate was associated with the production of ROS. Conclusion: These results suggest that butyrate generated by periodontal pathogens may be involved in the pathogenesis of periodontal diseases via the induction of ROS production and the impairment of cell growth, cell cycle progression and expression of cell cycle-related genes in GFs. These events are important in the initiation and prolongation of inflammatory processes in periodontal diseases.

Original languageEnglish
Pages (from-to)66-73
Number of pages8
JournalJournal of Periodontal Research
Volume48
Issue number1
DOIs
Publication statusPublished - Feb 2013

Fingerprint

Butyrates
Reactive Oxygen Species
Cell Cycle
Fibroblasts
Periodontal Diseases
Flow Cytometry
Western Blotting
G2 Phase Cell Cycle Checkpoints
cdc Genes
Polymerase Chain Reaction
Porphyromonas gingivalis
Butyric Acid
Trypan Blue
Volatile Fatty Acids
Propidium
Poisons
Periodontitis
Growth
Proteins
Cell Count

Keywords

  • Butyrate
  • Cell cycle
  • Periodontal pathogen
  • Periodontitis
  • Reactive oxygen species
  • Short-chain fatty acid

ASJC Scopus subject areas

  • Periodontics

Cite this

Butyrate induces reactive oxygen species production and affects cell cycle progression in human gingival fibroblasts. / Chang, M. C.; Tsai, Y. L.; Chen, Y. W.; Chan, C. P.; Huang, C. F.; Lan, W. C.; Lin, C. C.; Lan, W. H.; Jeng, J. H.

In: Journal of Periodontal Research, Vol. 48, No. 1, 02.2013, p. 66-73.

Research output: Contribution to journalArticle

Chang, M. C. ; Tsai, Y. L. ; Chen, Y. W. ; Chan, C. P. ; Huang, C. F. ; Lan, W. C. ; Lin, C. C. ; Lan, W. H. ; Jeng, J. H. / Butyrate induces reactive oxygen species production and affects cell cycle progression in human gingival fibroblasts. In: Journal of Periodontal Research. 2013 ; Vol. 48, No. 1. pp. 66-73.
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T1 - Butyrate induces reactive oxygen species production and affects cell cycle progression in human gingival fibroblasts

AU - Chang, M. C.

AU - Tsai, Y. L.

AU - Chen, Y. W.

AU - Chan, C. P.

AU - Huang, C. F.

AU - Lan, W. C.

AU - Lin, C. C.

AU - Lan, W. H.

AU - Jeng, J. H.

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N2 - Background and Objective: Short-chain fatty acids, such as butyric acid and propionic acid, are metabolic by-products generated by periodontal microflora such as Porphyromonas gingivalis, and contribute to the pathogenesis of periodontitis. However, the effects of butyrate on the biological activities of gingival fibroblasts (GFs) are not well elucidated. Material and Methods: Human GFs were exposed to various concentrations of butyrate (0.5-16mm) for 24h. Viable cells that excluded trypan blue were counted. Cell cycle distribution of GFs was analyzed by propidium iodide-staining flow cytometry. Cellular reactive oxygen species (ROS) production was measured by flow cytometry using 2',7'-dichlorofluorescein (DCF). Total RNA and protein lysates were isolated and subjected to RT-PCR using specific primers or to western blotting using specific antibodies, respectively. Results: Butyrate inhibited the growth of GFs, as indicated by a decrease in the number of viable cells. This event was associated with an induction of G0/G1 and G2/M cell cycle arrest by butyrate (4-16mm) in GFs. However, no marked apoptosis of GFs was noted in this experimental condition. Butyrate (>2mm) inhibited the expression of cdc2, cdc25C and cyclinB1 mRNAs and reduced the levels of Cdc2, Cdc25C and cyclinB1 proteins in GFs, as determined using RT-PCR and western blotting, respectively. This toxic effect of butyrate was associated with the production of ROS. Conclusion: These results suggest that butyrate generated by periodontal pathogens may be involved in the pathogenesis of periodontal diseases via the induction of ROS production and the impairment of cell growth, cell cycle progression and expression of cell cycle-related genes in GFs. These events are important in the initiation and prolongation of inflammatory processes in periodontal diseases.

AB - Background and Objective: Short-chain fatty acids, such as butyric acid and propionic acid, are metabolic by-products generated by periodontal microflora such as Porphyromonas gingivalis, and contribute to the pathogenesis of periodontitis. However, the effects of butyrate on the biological activities of gingival fibroblasts (GFs) are not well elucidated. Material and Methods: Human GFs were exposed to various concentrations of butyrate (0.5-16mm) for 24h. Viable cells that excluded trypan blue were counted. Cell cycle distribution of GFs was analyzed by propidium iodide-staining flow cytometry. Cellular reactive oxygen species (ROS) production was measured by flow cytometry using 2',7'-dichlorofluorescein (DCF). Total RNA and protein lysates were isolated and subjected to RT-PCR using specific primers or to western blotting using specific antibodies, respectively. Results: Butyrate inhibited the growth of GFs, as indicated by a decrease in the number of viable cells. This event was associated with an induction of G0/G1 and G2/M cell cycle arrest by butyrate (4-16mm) in GFs. However, no marked apoptosis of GFs was noted in this experimental condition. Butyrate (>2mm) inhibited the expression of cdc2, cdc25C and cyclinB1 mRNAs and reduced the levels of Cdc2, Cdc25C and cyclinB1 proteins in GFs, as determined using RT-PCR and western blotting, respectively. This toxic effect of butyrate was associated with the production of ROS. Conclusion: These results suggest that butyrate generated by periodontal pathogens may be involved in the pathogenesis of periodontal diseases via the induction of ROS production and the impairment of cell growth, cell cycle progression and expression of cell cycle-related genes in GFs. These events are important in the initiation and prolongation of inflammatory processes in periodontal diseases.

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KW - Periodontal pathogen

KW - Periodontitis

KW - Reactive oxygen species

KW - Short-chain fatty acid

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