The present study was conducted to screen candidate probiotic strains for anti-inflammatory activity. Initially, a nitric oxide (NO) assay was used to test selected candidate probiotic strains for anti-inflammatory activity in cultures of the murine macrophage cell line, RAW 264.7. Then, the in vitro probiotic properties of the strains, including bile tolerance, acid resistance, and growth in skim milk media, were investigated. We also performed an in vitro hydrophobicity test and an intestinal adhesion assay using Caenorhabditis elegans as a surrogate in vivo model. From our screening, we obtained 4 probiotic candidate lactic acid bacteria (LAB) strains based on their anti-inflammatory activity in lipopolysaccharide (LPS)-stimulated RAW 264.7 cell cultures and the results of the in vitro and in vivo probiotic property assessments. Molecular characterization using 16S rDNA sequencing analysis identified the 4 LAB strains as Lactobacillus plantarum. The selected L. plantarum strains (CAU1054, CAU1055, CAU1064, and CAU1106) were found to possess desirable in vitro and in vivo probiotic properties, and these strains are good candidates for further investigations in animal models and human clinical studies to elucidate the mechanisms underlying their anti-inflammatory activities.
(2014) Pyrosequencing analysis of microbiota reveals that lactic acid bacteria are dominant in Korean flat fish fermented food, gajami-sikhae, Bioscience, Biotechnology, and Biochemistry, 78:9, 1611-1618,
The results in this study improved our understanding of immunostimulating activity of PE-EPS and supported its potential treatment option as a natural immunostimulant.
We analyzed the bacterial community of Alaska pollock sikhae, a traditional Korean food made by natural fermentation with Alaska pollock, utilizing pyrosequencing. We fermented the Alaska pollock sikhae at two different temperatures (10 C and 20 C). Before fermentations, the bacterial community was varied. After fermentations, however, Lactobacillus sakei became dominant. The Alaska pollock sikhae sample before fermentations contained only 2% L. sakei, but the sample on day 6 of fermentation at 10 C comprised 74% L. sakei (90% at 20 C). In addition, we observed a reduction in the composition of unpreferred bacterial species for foods after fermentation. The composition of unpreferred bacterial species was more than 30% of total reads in samples before fermentation and decreased to less than 0.2% after fermentation. This result suggested that the fermentation of Alaska pollock sikhae can be beneficial for food safety. Alaska pollock sikhae might be a favorable habitat for L. sakei. Our study is the first report illustrating the alteration of the bacterial community of Alaska pollock sikhae during fermentation utilizing pyrosequencing analysis.Key words: Alaska pollock sikhae; bacterial community analysis; Lactobacillus sakei; pyrosequencing analysis IntroductionNumerous fermented foods are popular in Korea. Korean fermented foods were designed for long-term storage, to supplement various nutrients, and to provide featured flavors. Jeotgal and sikhae are both traditional fermented seafoods made for long-term storage (Rhee et al., 2011). While jeotgal contains a relatively high concentration of salt [generally 20 30% (w/w)], sikhae uses a low concentration of salt [ 7%] (Cha et al., 2004; Jung et al., 2013a, b;Guan et al., 2011;Lee et al., 2014). In contrast to jeotgal, sikhae contains grains that provide abundant carbon sources to microbes. Gajami-sikhae, Alaska pollock sikhae, and squid sikhae are the three most popular Korean sikhae. In addition to salt and fish (Alaska pollock), Alaska pollock sikhae is composed of cooked grains, white radish, red pepper powder, malt, garlic, and ginger. Alaska pollock sikhae is mainly divided into Kangwondo-and Kyoungsangdo-style recipes. While Kyoungsangdo-style recipes use dried Alaska pollock, undried fish are utilized for the Kangwondo-style recipes. Despite its importance as a Korean fermented food, there is limited research regarding sikhae and our understanding of the fermentation mechanisms of sikhae is largely incomplete.Novel bacterial strains were isolated from sikhae, mainly from gajami-sikhae (Kim et al., 2009;Jung et al., 2012a;Shin et al., 2011;Park et al., 2013Park et al., , 2014. Alishewanella jeotgali MS1 T was isolated from gajami-sikhae and its genome was fully sequenced (Kim et al., 2009;Jung et al., 2012a). Shin and co-workers isolated Leutobacter celer NAL101 T from gajami-sikhae (Shin et al., 2011). They suggested that the gajami-sikhae habitat might not be the optimal growth condition for the NAL101 T strain (Shin et al., 2011). Park and colleagues iso...
Fibrosis is induced by the excessive and abnormal deposition of extracellular matrix (ECM) with various growth factors in tissues. Transforming growth factor beta 1 (TGF-β1), plays a role in inducing apoptosis, modulates fibrosis, and ECM accumulation. Plasminogen activator inhibitor 1 (PAI-1) plays an important role in the development hepatic fibrosis. The overexpression of PAI-1 induces ECM accumulation, the main hallmark of chronic liver diseases. Death of hepatocytes is a characteristic feature of chronic liver disease due to various causes. The TGF-β1-mediated apoptotic pathway is regarded as a promising therapeutic target in hepatic fibrosis. Gamma-linolenic acid (GLA) is of special interest as it possesses anti-fibrosis, anti-inflammatory, and anti-cancer properties. However, the precise mechanism for GLA in chronic liver disease is not still clear. The aim of the present study was to determine whether GLA prevents hepatic PAI-1 expression and apoptosis through the inhibition of TGF-β1-mediated molecular mediators. GLA attenuated TGF-β1-stimulated PAI-1 expression, and inhibited PAI-1 promoter activity in AML12 cells. This effect was mediated by Smad3/4, the p38 pathways. We also found that GLA suppressed TGF-β1-induced apoptotic activation of the Bcl-2 family and caspase family of proteins, which resulted in the inhibition of poly-ADP-ribose polymerase 1 cleavage. GLA ameliorates the pro-fibrotic and pro-apoptotic effects of TGF-β1 in hepatocytes, suggesting GLA exerts a protective effect on hepatocytes and has a therapeutic potential for the treatment of chronic liver disease.
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