J Korean Acad Oral Health.  2018 Dec;42(4):224-228. 10.11149/jkaoh.2018.42.4.224.

Ethanol changes atpB gene expression and proton permeability in Streptococcus mutans

Affiliations
  • 1Department of Oral Biochemistry & Molecular Biology, School of Dentistry, Pusan National University, Yangsan, Korea. jsokang@pusan.ac.kr
  • 2Department of Preventive & Community Dentistry, School of Dentistry, Pusan National University, Yangsan, Korea.
  • 3BK21 PLUS Project, School of Dentistry, Pusan National University, Yangsan, Korea.

Abstract


OBJECTIVES
As a first step to study the anticaries effect of ethanol alone, we investigated the effects of ethanol on the expression levels of the atpB gene and proton permeability of Streptococcus mutans in suspension cultures.
METHODS
S. mutans UA159 was grown in brain heart infusion medium at either pH 4.8 or 6.8. The total extracted RNA was reverse-transcribed into cDNA using a Superscriptâ„¢ First-Strand Synthesis System. The resulting cDNA and negative controls were amplified by ABI PRISM 7700 real-time PCR system with SYBR Green PCR Master Mix. For proton flux assay, bacterial suspensions were titrated to pH 4.6 with 0.5 M HCl, and then additional 0.5 M HCl was added to decrease the pH values by approximately 0.4 units. The subsequent increase in pH was monitored using a glass electrode. Ten percent (v/v) butanol was added to the suspensions at 80 min to disrupt the cell membrane.
RESULTS
In a concentration-dependent manner, ethanol alone not only decreased the growth rate of S. mutans and the expression of the atpB gene but also increased the proton permeability at both pH 4.8 and 6.8.
CONCLUSIONS
These findings suggest that ethanol has the potential for an anticaries ingredient. We believe that ethanol may be used together with fluoride and/or other cariostatic agents in order to develop better anticaries toothpastes and/or mouthrinses.

Keyword

Ethanol; atpB gene expression; Proton permeability; Streptococcus mutans

MeSH Terms

Brain
Cariostatic Agents
Cell Membrane
DNA, Complementary
Electrodes
Ethanol*
Fluorides
Gene Expression*
Glass
Heart
Hydrogen-Ion Concentration
Permeability*
Polymerase Chain Reaction
Protons*
Real-Time Polymerase Chain Reaction
RNA
Streptococcus mutans*
Streptococcus*
Suspensions
Toothpastes
Cariostatic Agents
DNA, Complementary
Ethanol
Fluorides
Protons
RNA
Suspensions
Toothpastes

Figure

  • Fig. 1 Growth curves of S. mutans UA159. The growth rates (c) were calculated as c=Δlog2(OD420)/Δt where OD420 is the optical density at 420 nm, and t is time. Data are reported as the mean±SEM of triplicate analysis of three preparations.

  • Fig. 2 The relative mRNA levels for the atpB gene of S. mutans UA159. The relative values were obtained by normalizing the atpB mRNA with expression of 16S rRNA gene. The Data are reported as mean±SEM of triplicate analysis of three preparations. *P<0.05, **P<0.01, ***P<0.001.

  • Fig. 3 Effects of ethanol on the proton permeability in S. mutans UA159. Data are reported as the mean±SEM of triplicate analysis of three preparations.


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