Moon, Ji Eun
(Department of Food and Biotechnology, Korea University)
,
Heo, Wan
(Institutes of Natural Sciences, Korea University)
,
Lee, Sang Hoon
(Department of Food and Biotechnology, Korea University)
,
Lee, Suk Hee
(Department of Molecular Medicine, Cell and Matrix Research Institute, Kyungpook National University School of Medicine)
,
Lee, Hong Gu
(Department of Animal Science and Technology, College of Animal Bioscience and Technology, Konkuk University)
,
Lee, Jin Hyup
(Department of Food and Biotechnology, Korea University)
,
Kim, Young Jun
(Department of Food and Biotechnology, Korea University)
Saccharomyces boulardii is the only probiotic yeast with US Food and Drug Administration approval. It is routinely used to prevent or treat acute diarrhea and other gastrointestinal disorders, including the antibiotic-associated diarrhea caused by Clostridium difficile infections. The formation of r...
Saccharomyces boulardii is the only probiotic yeast with US Food and Drug Administration approval. It is routinely used to prevent or treat acute diarrhea and other gastrointestinal disorders, including the antibiotic-associated diarrhea caused by Clostridium difficile infections. The formation of reactive oxygen species (ROS), specifically H2O2 during normal aerobic metabolism, contributes to programmed cell death and represents a risk to the viability of the probiotic microbe. Moreover, a loss of viability reduces the efficacy of the probiotic treatment. Therefore, inhibiting the accumulation of ROS in the oxidant environment could improve the viability of the probiotic yeast and lead to more efficacious treatment. Here, we provide evidence that supplementation with a non-reducing disaccharide, namely trehalose, enhanced the viability of S. boulardii exposed to an oxidative environment by preventing metacaspase YCA1-mediated programmed cell death through inhibition of intracellular ROS production. Our results suggest that supplementation with S. boulardii together with trehalose could increase the viability of the organism, and thus improve its effectiveness as a probiotic and as a treatment for acute diarrhea and other gastrointestinal disorders.
Saccharomyces boulardii is the only probiotic yeast with US Food and Drug Administration approval. It is routinely used to prevent or treat acute diarrhea and other gastrointestinal disorders, including the antibiotic-associated diarrhea caused by Clostridium difficile infections. The formation of reactive oxygen species (ROS), specifically H2O2 during normal aerobic metabolism, contributes to programmed cell death and represents a risk to the viability of the probiotic microbe. Moreover, a loss of viability reduces the efficacy of the probiotic treatment. Therefore, inhibiting the accumulation of ROS in the oxidant environment could improve the viability of the probiotic yeast and lead to more efficacious treatment. Here, we provide evidence that supplementation with a non-reducing disaccharide, namely trehalose, enhanced the viability of S. boulardii exposed to an oxidative environment by preventing metacaspase YCA1-mediated programmed cell death through inhibition of intracellular ROS production. Our results suggest that supplementation with S. boulardii together with trehalose could increase the viability of the organism, and thus improve its effectiveness as a probiotic and as a treatment for acute diarrhea and other gastrointestinal disorders.
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가설 설정
Recently, a protective effect of polyphenols against the toxic effects of ROS in yeast was reported [21-23]. Thus, it can be hypothesized that under the circumstances of oxidative stress, probiotic yeast might be protected using natural or synthetic antioxidants that would scavenge the ROS, and lead to increased cell viability. In the present study, the non-reducing disaccharide, trehalose (α-D-glucopyranosyl-1,1-α-D-glucopyranoside), was selected as an antioxidant, since it has been reported to possess a strong ROS-scavenging property [17, 24].
제안 방법
The identification of S. boulardii was performed by the PCR-based amplification of the internal transcribed spacer (ITS) region between the rDNA subunits, including the gene that encodes 5.8S rRNA, using primers for ITS2 (forward primer: 5’-AGGTTTTACCAACTGCGGCT-3’; reverse primer: 5’-TCGCCTAGACGCTCTCTTCT-3’) and microsatellite loci using primers for YLR177w (forward primer: 5’-CTTAAA CAACAGCTCCCAAA-3’; reverse primer: 5’-ATGAATCAGCGC ATCAGAAAT-3’).
boulardii. Therefore, to investigate the molecular mechanisms by which trehalose protected against cytotoxicity in S. boulardii exposed to oxidative stress, H2O2-exposed cells, with or without trehalose pretreatment were tested for PCD using a TUNEL assay. As shown in Fig.
2A), suggesting that trehalose protected them against oxidant-induced cell death. To further evaluate the intracellular perturbations contributing to the growth inhibition of the cells observed after exposure to H2O2, we examined their metabolic activity using the FUN-1 stain. In cells stained with FUN-1, a higher red-to-green fluorescence ratio indicates a higher metabolic activity [27].
대상 데이터
All experiments were performed with S. boulardii (CNCM I-1079) obtained from LALLEMAND Inc. (Canada). The yeast was grown at 30°C in a standard liquid yeast extract peptone dextrose (YPD) broth (BD Difco, DF0427-17-6) on a rotary shaker at 200 rpm or on YPD agar containing 2% agar.
데이터처리
Statistical analyses were performed by two-tailed t-tests. Results are shown as mean ± standard deviation.
성능/효과
Taken together, our results proved that the extent of PCD in S. boulardii cells exposed to oxidative stress was significantly reduced by treating the organism with trehalose, which was reflected by increased cell survival under the oxidative environment.
참고문헌 (35)
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