Document Detail


Butyrate, a bacterial metabolite, induces apoptosis and autophagic cell death in gingival epithelial cells.
MedLine Citation:
PMID:  20546110     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND AND OBJECTIVE: Butyrate is produced by some types of anaerobic periodontal bacteria. Millimolar concentrations of butyrate are found in mature dental plaque from periodontitis patients. Although butyrate reportedly has a variety of effects in many mammalian cells, its effect on gingival epithelial cells is not well known. In this study, we investigated the effect of butyrate on gingival epithelial Ca9-22 cell death.
MATERIAL AND METHODS: Death of Ca9-22 cells was assessed after treating the cells with or without butyrate. A SYTOX Green dye, which exhibits strong green fluorescence once it enters dead cells through ruptured cell membranes, was used for cell death detection. Phosphatidylserine redistribution was measured using fluorescein isothiocyanate-labeled annexin V. The activity of caspase-3 was measured as the amount of cleaved substrate peptide. Anti-apoptotic bcl-2 mRNA expression was measured using real-time RT-PCR. Western blotting and fluoromicroscopic analysis with anti-microtubule-associated protein 1 light chain 3 (LC3) antibodies were performed for detection of autophagy.
RESULTS: Stimulation with millimolar concentrations of butyrate for 48 h induced Ca9-22 cell death. The stimulation also caused increased caspase-3 activity, phosphatidylserine redistribution and bcl-2 down-regulation, suggesting butyrate-induced apoptosis. However, the pan-caspase inhibitor, Z-VAD-FMK, did not inhibit cell death completely. This implies the existence of other types of cell death. In addition, markers of autophagy, namely, the conversion of LC3-I to LC3-II and increased LC3 accumulation, were observed. Moreover, inhibition of autophagy by 3-methyladenine suppressed the butyrate-induced cell death, suggesting that butyrate could induce cell death through autophagy.
CONCLUSION: These data suggest that butyrate induces apoptosis and autophagic cell death.
Authors:
H Tsuda; K Ochiai; N Suzuki; K Otsuka
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-06-10
Journal Detail:
Title:  Journal of periodontal research     Volume:  45     ISSN:  1600-0765     ISO Abbreviation:  J. Periodont. Res.     Publication Date:  2010 Oct 
Date Detail:
Created Date:  2010-08-24     Completed Date:  2010-12-06     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0055107     Medline TA:  J Periodontal Res     Country:  Denmark    
Other Details:
Languages:  eng     Pagination:  626-34     Citation Subset:  D; IM    
Copyright Information:
(c) 2010 John Wiley & Sons A/S.
Affiliation:
Department of Biochemistry, Nihon University School of Dentistry, Tokyo, Japan. tsuda-h@dent.nihon-u.ac.jp
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MeSH Terms
Descriptor/Qualifier:
Annexin A5 / diagnostic use
Apoptosis*
Autophagy*
Bacteria, Anaerobic / metabolism
Butyrates / pharmacology*
Caspase 3 / antagonists & inhibitors,  metabolism
Cell Line, Tumor
Down-Regulation
Epithelial Cells / drug effects*
Fluorescein-5-isothiocyanate / diagnostic use
Fluorescent Dyes / diagnostic use
Gingiva / cytology,  drug effects*
Humans
Organic Chemicals / diagnostic use
Phosphatidylserines / metabolism
Reverse Transcriptase Polymerase Chain Reaction
bcl-Associated Death Protein / biosynthesis
Chemical
Reg. No./Substance:
0/Annexin A5; 0/Butyrates; 0/Fluorescent Dyes; 0/Organic Chemicals; 0/Phosphatidylserines; 0/SYTOX Green; 0/bcl-Associated Death Protein; 3326-32-7/Fluorescein-5-isothiocyanate; EC 3.4.22.-/Caspase 3

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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