• Title/Summary/Keyword: Caco-2 cells

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Selection and immunomodulatory evaluation of lactic acid bacteria suitable for use as canine probiotics (개 생균제 사용에 적합한 유산균주의 선발 및 면역활성 평가)

  • Park, Su-Min;Park, Ho-Eun;Lee, Wan-Kyu
    • Korean Journal of Veterinary Research
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    • v.55 no.2
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    • pp.81-88
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    • 2015
  • This study was conducted to isolate lactic acid bacteria (LAB) from dog intestine and identify potential probiotic strains for canine use. One hundred and one LAB were isolated from feces of 20 healthy dogs. Acid, bile, and heat resistance along with adherence to Caco-2 cells and antimicrobial activity against pathogens were examined. To analyze immunomodulative effects, the production of nitric oxide (NO), TNF-${\alpha}$, and IL-$1{\beta}$ was measured using RAW 264.7 macrophages. Additionally, RAW BLUE cells were used to evaluate nuclear factor-${\kappa}B$ (NF-${\kappa}B$) generation. Ultimately, three strains were selected as canine probiotics and identified as Lactobacillus reuteri L10, Enterococcus faecium S33, and Bifidobacterium longum B3 by 16S rRNA sequence analysis. The L10 and S33 strains showed tolerance to pH 2.5 for 2 h, 1.0% Oxgall for 2 h, and $60^{\circ}C$ for 5 min. These strains also had strong antimicrobial activity against Escherichia coli KCTC 1682, Salmonella Enteritidis KCCM 12021, Staphylococcus aureus KCTC 1621, and Listeria monocytogenes KCTC 3569. All three strains exerted better immunomodulatory effects than Lactobacillus rhamnosus GG (LGG), a well-known commercial immunomodulatory strain, based on NO, NF-${\kappa}B$, IL-$1{\beta}$, and TNF-${\alpha}$ production. These results suggested that the three selected strains could serve as canine probiotics.

Anti-Obesity Potential through Regulation of Carbohydrate Uptake and Gene Expression in Intestinal Epithelial Cells by the Probiotic Lactiplantibacillus plantarum MGEL20154 from Fermented Food

  • So Young Park;Jin Won Choi;Dong Nyoung Oh;Eun Ji Lee;Dong Pil Kim;Sun Jay Yoon;Won Je Jang;Sang Jun Han;Seungjun Lee;Jong Min Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.5
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    • pp.621-633
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    • 2023
  • We investigated the probiotic characteristics and anti-obesity effect of Lactiplantibacillus plantarum MGEL20154, a strain that possesses excellent intestinal adhesion and viability. The in vitro properties, e.g., gastrointestinal (GI) resistance, adhesion, and enzyme activity, demonstrated that MGEL20154 is a potential probiotic candidate. Oral administration of MGEL20154 to diet-induced obese C57BL/6J mice for 8 weeks resulted in a feed efficacy decrease by 44.7% compared to that of the high-fat diet (HFD) group. The reduction rate of weight gain was about 48.5% in the HFD+MGEL20154 group compared to that of the HFD group after 8 weeks, and the epididymal fat pad was also reduced in size by 25.2%. In addition, the upregulation of the zo-1, pparα, and erk2, and downregulation of the nf-κb and glut2 genes in Caco-2 cells by MGEL20154 were observed. Therefore, we propose that the anti-obesity effect of the strain is exerted by inhibiting carbohydrate absorption and regulating gene expression in the intestine.

Antioxidant, Antibacterial and Anti-inflammatory Effects of Stachys sieboldii Extract (초석잠 추출물의 항산화, 항균 및 항염 활성)

  • Lee, Jeong Ho
    • Korean Journal of Plant Resources
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    • v.34 no.5
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    • pp.420-432
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    • 2021
  • This study was conducted to evaluate antioxidant, antibacterial, anti-inflammatory, and digestive enzyme activity in water extract (SAW) and 60% ethanol extract (SAE) from Stachys sieboldii. As the treatment concentration of each extract S. sieboldii extract increased, antibacterial and antioxidant activity increased. The total polyphenol and total flavonoid contents of SAW were 106.25 ± 0.94 mgGAE/g, 24.4 ± 0.24 mgQE/g and SAE were 124.61 ± 1.11 mgGAE/g, 45.2 ± 3.52 mgQE/g, respectively. The 400 ㎍/mL of SAW and SAE performed more than 53% protective effects against oxidative stress in HepG2 cell lines. All extracts were not showed cytotoxicity to RAW 264.7 cell line at 100 ㎍/mL. NO production was reduced to 44.3 ± 1.4% for SAW and 45.1 ± 1.0% for SAE at a concentration of 100 ㎍/mL. The production of inflammatory cytokines each TNF-α, IL-1β, and IL-6 was inhibited in a concentration-dependent. S. sieboldii extract did not showed Caco-2 cells cytotoxicity and inhibited NO production in concentration-dependent. As the concentration of the S. sieboldii extract increased of α-amylase and protease enzymes activity, which are digestive enzyme. As a result of the experiment, it is judged that it can be used as basic data for the development of health food using S. sieboldii.

Immunomodulatory Effects of Bifidobacterium spp. and Use of Bifidobacterium breve and Bifidobacterium longum on Acute Diarrhea in Children

  • Choi, Yae Jin;Shin, Seon-Hee;Shin, Hea Soon
    • Journal of Microbiology and Biotechnology
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    • v.32 no.9
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    • pp.1186-1194
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    • 2022
  • The intake of probiotic lactic acid bacteria not only promotes digestion through the microbiome regulated host intestinal metabolism but also improves diseases such as irritable bowel syndrome and inflammatory bowel disease, and suppresses pathogenic harmful bacteria. This investigation aimed to evaluate the immunomodulatory effects in intestinal epithelial cells and to study the clinical efficacy of the selected the Bifidobacterium breve and Bifidobacterium longum groups. The physiological and biochemical properties were characterized, and immunomodulatory activity was measured against pathogenic bacteria. In order to find out the mechanism of inflammatory action of the eight viable and sonicated Bifidobacterium spp., we tried to confirm the changes in the pro-inflammatory cytokines (TNF-α, interleukin (IL)-6, IL-12) and anti-inflammatory cytokine (IL-10), and chemokines, (monocyte chemoattractant protein-1, IL-8) and inflammatory enzymatic mediator (nitric oxide) against Enterococcus faecalis ATCC 29212 infection in Caco-2 cells and RAW 264.7 cells. The clinical efficacy of the selected B. breve and B. longum group was studied as a probiotic adjuvant for acute diarrhea in children by oral administration. The results showed significant immunomodulatory effects on the expression levels of TNF-α, IL-6, IL-12, MCP-1, IL-8 and NO, in sonicated Bifidobacterium extracts and viable bifidobacteria. Moreover, each of the Bifidobacterium strains was found to react more specifically to different cytokines. However, treatment with sonicated Bifidobacterium extracts showed a more significant effect compared to treatment with the viable bacteria. We suggest that probiotics functions should be subdivided according to individual characteristics, and that personalized probiotics should be designed to address individual applications.

Ginsenoside Rg3 reduces the adhesion, invasion, and intracellular survival of Salmonella enterica serovar Typhimurium

  • Mechesso, Abraham F.;Quah, Yixian;Park, Seung-Chun
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.75-85
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    • 2021
  • Background: Invasive infections due to foodborne pathogens, including Salmonella enterica serovar Typhimurium, are prevalent and life-threatening. This study aimed to evaluate the effects of ginsenoside Rg3 (Rg3) on the adhesion, invasion, and intracellular survival of S. Typhimurium. Methods: The impacts of Rg3 on bacterial growth and host cell viability were determined using the time kill and the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assays, respectively. Gentamicin assay and confocal microscopic examination were undertaken to determine the effects of Rg3 on the adhesive and invasive abilities of S. Typhimurium to Caco-2 and RAW 264.7 cells. Quantitative reverse transcription polymerase chain reaction was performed to assess the expression of genes correlated with the adhesion, invasion, and virulence of S. Typhimurium. Results: Subinhibitory concentrations of Rg3 significantly reduced (p < 0.05) the adhesion, invasion, and intracellular survival of S. Typhimurium. Rg3 considerably reduced (p < 0.05) the bacterial motility as well as the release of nitrite from infected macrophages in a concentration-dependent manner. The expression of genes related to the adhesion, invasion, quorum sensing, and virulence of S. Typhimurium including cheY, hilA, OmpD, PrgK, rsgE, SdiA, and SipB was significantly reduced after Rg3 treatment. Besides, the compound downregulated rac-1 and Cdc-42 that are essential for actin remodeling and membrane ruffling, thereby facilitating Salmonella entry into host cells. This report is the first to describe the effects of Rg3 on "trigger" entry mechanism and intracellular survival S. Typhimurium. Conclusion: Rg3 could be considered as a supplement agent to prevent S. Typhimurium infection.

Probiotic Properties of Lactobacillus strains Isolated from Kimchi (김치로부터 분리된 Lactobacillus strains의 probiotic 특성)

  • Choi, Hye Jung;Lim, Bo Ram;Kim, Dong Wan;Kwon, Gi-Seok;Joo, Woo Hong
    • Journal of Life Science
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    • v.24 no.11
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    • pp.1231-1237
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    • 2014
  • The objective of this study was to evaluate the safety and functional properties of four potential probiotic strains isolated from Kimchi, traditional Korean fermented vegetables. Based on being higher tolerance to bile salts and showing higher acid resistance or hydrophobic properties, one Lactobacillus arizonensis strain (BCNU 9032) and three L. brevis strains (BCNU 9037, BCNU 9098 and BCNU 9101) were selected in the screening experiment. All strains can survived up to 99% after 3h culture in pH 2.5 and resistant to 1% bile salts. These strains also showed good antimicrobial activities against a number of food borne pathogens, especially against Escherichia coli and Shigella sonnei. The ability to lower cholesterol levels of L. arizonensis BCNU 9032 and L. brevis 9037 were demonstrated by bile salt hydrolytic activity and cholesterol assimilation tests. Moreover, L. brevis BCNU 9098 and BCNU 9101 showed higher adherence to Caco-2 cells (12.76 and 11.86%, respectively) than Lactobacillus rhamnosus GG, a commercial probiotic strain used worldwide. The results suggest that these strains could be used as probiotics.

Characterization of Lactobacillus plantarum strains isolated from black raspberry and their effect on BALB/c mice gut microbiota

  • Choi, Hye Ran;Chung, Yi Hyung;Yuk, Hyun-Gyun;Lee, Hyunki;Jang, Han Su;Kim, Yosum;Shin, Daekeun
    • Food Science and Biotechnology
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    • v.27 no.6
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    • pp.1747-1754
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    • 2018
  • The objective of this study was to evaluate probiotic effects of two Lactobacillus plantarum strains (GBL16 and 17) isolated from black raspberry. Results revealed that the number of GBL16 was gradually decreased as bile salt concentration was increased from 0.3 to 1%. However, GBL17 did not show any difference when GBL17 was applied to 1% bile salt, and it indicates that GBL17 is more tolerant to bile salt than GBL16. GBL17 exhibited higher heat resistance and adhesion ability to Caco-2 cells than GBL16. Regarding gut microbiome, no significant change in the number of total bacteria in intestines of mice after treatment with GBLs was determined. However, the combination of GBL16 and GBL17 significantly increased the number of total bacteria in intestines of mice after they were orally administered. Therefore, the results suggest that both GBL16 and 17 strains could be one of major probiotics that can improve human gut health.

The Transport of Organic Cations in the Small Intestine: Current Knowledge and Emerging Concepts

  • Kim, Moon-Kyoung;Shim, Chang-Koo
    • Archives of Pharmacal Research
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    • v.29 no.7
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    • pp.605-616
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    • 2006
  • A wide variety of drugs and endogenous bioactive amines are organic cations (OCs). Approximately 40% of all conventional drugs on the market are OCs. Thus, the transport of xenobiotics or endogenous OCs in the body has been a subject of considerable interest, since the discovery and cloning of a family of OC transporters, referred to as organic cation transporter (OCTs), and a new subfamily of OCTs, OCTNs, leading to the functional characterization of these transporters in various systems including oocytes and some cell lines. Organic cation transporters are critical in drug absorption, targeting, and disposition of a drug. In this review, the recent advances in the characterization of organic cation transporters and their distribution in the small intestine are discussed. The results of the in vitro transport studies of various OCs in the small intestine using techniques such as isolated brush-border membrane vesicles, Ussing chamber systems and Caco-2 cells are discussed, and in vivo knock-out animal studies are summarized. Such information is essential for predicting pharmacokinetics and pharmacodynamics and in the design and development of new cationic drugs. An understanding of the mechanisms that control the intestinal transport of OCs will clearly aid achieving desirable clinical outcomes.

Probiotic properties and safety assessment of lactic acid bacteria isolated from salt-fermented anchovy (멸치 젓갈로부터 분리된 젖산세균의 프로바이오틱 특성 및 안전성 평가)

  • Lim, Eun-Seo;Kim, Young-Mog;Lee, Eun-Woo
    • Korean Journal of Food Science and Technology
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    • v.48 no.4
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    • pp.306-316
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    • 2016
  • This study was conducted to evaluate the functional characteristics and safety properties of lactic acid bacteria isolated from salt-fermented anchovy, a putative probiotic candidate. The following isolates were identified by biochemical profiles, carbohydrate fermentation patterns, and 16S rRNA sequencing: Enterococcus faecium AJ06, Leuconostoc mesenteroides AJ13, Pediococcus halophilus AJ22, Lactobacillus sakei AJ29, and Pediococcus pentosaceus AJ35. The strains AJ06, AJ22, AJ29 exhibited high tolerance to simulated gastric and intestinal juices and were able to produce bile salt hydrolase on MRS agar plates supplemented with taurocholic acid and/or taurodeoxycholic acid. The strains AJ22 and AJ29, which demonstrated high adherence to Caco-2 cells and resistance to various antibiotics, effectively inhibited the growth of food-borne pathogens by the production of antimicrobial substances. These strains did not show ${\alpha}-$ or ${\beta}$-haemolysis on blood agar. Furthermore, biogenic amines in MRS broth containing the precursor amino acids were not mutagenic in Salmonella Typhimurium TA98 and TA100.

Heat shock protein 90β inhibits apoptosis of intestinal epithelial cells induced by hypoxia through stabilizing phosphorylated Akt

  • Zhang, Shuai;Sun, Yong;Yuan, Zhiqiang;Li, Ying;Li, Xiaolu;Gong, Zhenyu;Peng, Yizhi
    • BMB Reports
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    • v.46 no.1
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    • pp.47-52
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    • 2013
  • Intestinal epithelial cell (IEC) apoptosis induced by hypoxia compromise intestinal epithelium barrier function. Both Akt and Hsp90 have cytoprotective function. However, the specific role of Akt and $Hsp90{\beta}$ in IEC apoptosis induced by hypoxia has not been explored. We confirmed that hypoxia-induced apoptosis was reduced by $Hsp90{\beta}$ overexpression but enhanced by decreasing $Hsp90{\beta}$ expression. $Hsp90{\beta}$ overexpression enhanced BAD phosphorylation and thus reduced mitochondrial release of cytochrome C. Reducing $Hsp90{\beta}$ expression had opposite effects. The protective effect of $Hsp90{\beta}$ against apoptosis was negated by LY294002, an Akt inhibitor. Further study showed that Akt phosphorylation was enhanced by $Hsp90{\beta}$, which was not due to the activation of upstream PI3K and PDK1 but because of stabilization of pAkt via direct interaction between $Hsp90{\beta}$ and pAkt. These results demonstrate that $Hsp90{\beta}$ may play a significant role in protecting IECs from hypoxia-induced apoptosis via stabilizing pAkt to phosphorylate BAD and reduce cytochrome C release.