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Butyrate, a bacterial metabolite, induces apoptosis and autophagic cell death in gingival epithelial cells.

Abstract：

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.

journal_name

J Periodontal Res

journal_title

Journal of periodontal research

authors

Tsuda H,Ochiai K,Suzuki N,Otsuka K

doi

10.1111/j.1600-0765.2010.01277.x

subject

Has Abstract

pub_date

2010-10-01 00:00:00

pages

626-34

issue

eissn

0022-3484

issn

1600-0765

pii

JRE1277

journal_volume

pub_type

杂志文章

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Butyrate, a bacterial metabolite, induces apoptosis and autophagic cell death in gingival epithelial cells.