• Title/Summary/Keyword: HT29 cells

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The Effects of Chelidonium majus on NO and $TNF-{\alpha}$ Production in Macrophages (백굴채가 대식세포의 NO 및 $TNF-{\alpha}$ 생성에 미치는 영향)

  • 김홍준;문석재;김동웅;문구;원경숙;윤준철;김유경;원진희
    • The Journal of Korean Medicine
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    • v.24 no.2
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    • pp.138-147
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    • 2003
  • Objectives : In this study, we investigated the mechanism by which Chelidonium majus (CM) regulates nitric oxide (NO) production. Methods : Using mouse peritoneal macrophages, the mechanism by which CM regulates NO or tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ production was examined. NO release was measured by the Griess method. $TNF-{\alpha}$ production was measured by the ELISA method. The protein extracts were prepared and samples were analyzed for the inducible NOS(iNOS) expression and nuclear factor kappa $B(NF-{\kappa}B)$ activation by Western blotting. Results : When CM was used in combination with recombinant $interferon-{\gamma}{\;}(rIFN-{\gamma})$, there was a marked cooperative induction of NO production. CM had an effect on NO production by itself. The expression of the iNOS gene was increased in $rIFN-{\gamma}$ plus CM-stimulated peritoneal macrophages and almost completely inhibited by pre-treatment with pyrrolidine dithiocarbamate (PDTC), an inhibitor of $NF-{\kappa}B$. The $NF-{\kappa}B$ activation was increased in rIFN-{\gamma} plus CM-induced peritoneal macrophages. The increased production of NO from $rIFN-{\gamma}$ plus CM-stimulated peritoneal rnacrophages was decreased by the treatment with $N^{G}-monomethyl-{_L}-arginine{\;}(N^{G}MMA){\;}N^{\alpha}-Tosyl-Phe$ chloromethyl ketone (TPCK) , and was almost completely inhibited by pre-treatment with PDTC. Furthermore, treatment with CM alone or rIFN-{\gamma} plus CM in peritoneal macrophages caused a significant increase in $TNF-{\alpha}$ production. PDTC decreased CM-induced $TNF-{\alpha}$ production significantly. After CM treatment in HT-29 or AGS cells, cell viability decreased. Conclusions : These findings demonstrate that CM increases the production of NO and $TNF-{\alpha}{\;}by{\;}rIFN-{\gamma}-primed$ macrophages and suggest that NF-B plays a critical role in mediating these effects of CM.

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Detection of Ref-1 (Redox factor-1) Interacting Protein Using the Yeast Two-hybrid System (Yeast two-hybrid system을 이용한 Ref-1 (redox factor-1) 결합 단백질의 분리 및 동정)

  • 이수복;김규원;배문경;배명호;정주원;안미영;김영진
    • Journal of Life Science
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    • v.14 no.1
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    • pp.26-31
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    • 2004
  • Redox factor-1 (Ref-1), known as a redox regulator, controls the DNA binding of AP-1 and is activated in HT29 colon cancer cells by hypoxia in vitro. REF-1 also increases tile DNA binding affinity of Hypoxia-inducible Factor-lalpha$ (HIF-lalpha$), HIF-like Factor (HLF) and early growth response-1 (Egr-1) which induce expression of the genes involved in angiogenesis, so that we speculate that REF-1 may play a role in hypoxia-induced angiogenesis. In this research we tried to detect novel proteins interacting with REF-1 using Yeast two-hybrid system using full-length REF-1 cDNA as bait. As result of such screening we detected 3 positive clones. DNA sequencing and GeneBank search revealed that one of the clones contained the same sequences as M.musculus cDNA for tioredoxin.

Influence of Extraction Method on Quality and Functionality of Broccoli Juice

  • Lee, Sung Gyu;Kim, Jin-Hee;Son, Min-Jung;Lee, Eun-Ju;Park, Woo-Dong;Kim, Jong-Boo;Lee, Sam-Pin;Lee, In-Seon
    • Preventive Nutrition and Food Science
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    • v.18 no.2
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    • pp.133-138
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    • 2013
  • This study was performed to compare the quality and functionality of broccoli juice as affected by extraction method. Broccoli juice was extracted using method I (NUC Kuvings silent juicer), method II (NUC centrifugal juicer), and method III (NUC mixer), and the quality properties of the broccoli juices were analyzed using three different methods. Additionally, the antioxidative, anticancer, and anti-hyperglycemic activities of broccoli juice prepared by the three different methods were investigated in vitro. The broccoli juice made by method I contained the highest polyphenol and flavonoid contents at 1,226.24 mg/L and 1,018.32 mg/L, respectively. Particularly, broccoli juice prepared by method I showed higher DPPH and ABTS radical scavenging activities than those of the other samples. Additionally, broccoli juice made by method I showed the highest growth inhibitory effects against HeLa, A549, AGS, and HT-29 cancer cells. Broccoli juice prepared by method I had the highest ${\alpha}$-glucosidase inhibitory effects. These results indicate that there are important differences in chemical and functional qualities between juice extraction techniques.

Isolation and characterization of cholesterol-lowering lactic acid bacteria from kimchi (김치에서 분리된 콜레스테롤 감소능을 가진 젖산세균의 특성)

  • Park, Hong-Yeop;Park, Seul-Ki;Kim, Bo-Geum;Ryu, Dae-Gyu;Lim, Eun-Seo;Kim, Young-Mog
    • Korean Journal of Food Science and Technology
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    • v.49 no.4
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    • pp.377-382
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    • 2017
  • The objective of this study was to isolate and characterize lactic acid bacteria (LAB) exhibiting cholesterol-lowering activity from the Korean traditional fermented food, kimchi. The previously isolated LAB strains were assessed for cholesterol-lowering efficacy in the presence of 0.1% cholesterol. All LAB strains tested in this study were able to assimilate cholesterol at varying levels, ranging from 35.0 to 99.4%. Among them, the Lactobacillus plantarum FMB 31 strain exhibited the highest cholesterol-lowering effect with 99.4% cholesterol removal efficiency. The strain was stable in the presence of acid, bile, and salt stress, and showed high adherence on HT-29 cells, a human colon line. In addition, the LAB strain showed no pathogenic properties such as the production of hemolysin and biogenic amines. Thus, this study suggests that the L. plantarum FMB 31 strain isolated from kimchi can be a potential source of probiotic products with strong cholesterol-lowering effect.

Comparative Genomic Analysis of Lactobacillus rhamnosus BFE5264, a Probiotic Strain Isolated from Traditional Maasai Fermented Milk

  • Jeong, Haeyoung;Choi, Sanghaeng;Park, Gun-Seok;Ji, Yosep;Park, Soyoung;Holzapfel, Wilhelm Heinrich;Mathara, Julius Maina;Kang, Jihee
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.25-33
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    • 2019
  • Lactobacillus rhamnosus BFE5264, isolated from a Maasai fermented milk product ("kule naoto"), was previously shown to exhibit bile acid resistance, cholesterol assimilation, and adhesion to HT29-MTX cells in vitro. In this study, we re-annotated and analyzed the previously reported complete genome sequence of strain BFE5264. The genome consists of a circular chromosome of 3,086,152 bp and a putative plasmid, which is the largest one identified among L. rhamnosus strains. Among the 2,883 predicted protein-coding genes, those with carbohydrate-related functions were the most abundant. Genome analysis of strain BFE5264 revealed two consecutive CRISPR regions and no known virulence factors or antimicrobial resistance genes. In addition, previously known highly variable regions in the genomes of L. rhamnosus strains were also evident in strain BFE5264. Pairwise comparison with the most studied probiotic strain L. rhamnosus GG revealed strain BFE5264-specific deletions, probably due to insertion sequence-mediated recombination. The latter was associated with loss of the spaCBA pilin gene cluster and exopolysaccharide biosynthetic genes. Comparative genomic analysis of the sequences from all available L. rhamnosus strains revealed that they were clustered into two groups, being within the same species boundary based on the average nucleotide identities. Strain BFE5264 had a sister group relationship with the group that contained strain GG, but neither ANI-based hierarchical clustering nor core-gene-based phylogenetic tree construction showed a clear distinctive pattern associated with the isolation source, implying that the genotype alone cannot account for their ecological niches. These results provide insights into the probiotic mechanisms of strain BFE5264 at the genomic level.

Actinidia arguta Sprout as a Natural Antioxidant: Ameliorating Effect on Lipopolysaccharide-Induced Cognitive Impairment

  • Kang, Jeong Eun;Park, Seon Kyeong;Kang, Jin Yong;Kim, Jong Min;Kwon, Bong Seok;Park, Sang Hyun;Lee, Chang Jun;Yoo, Seul Ki;Heo, Ho Jin
    • Journal of Microbiology and Biotechnology
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    • v.31 no.1
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    • pp.51-62
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    • 2021
  • Here, we investigated the prebiotic and antioxidant effects of Actinidia arguta sprout water extract (AASWE) on lipopolysaccharide (LPS)-induced cognitive deficit mice. AASWE increased viable cell count, titratable acidity, and acetic acid production in Lactobacillus reuteri strain and showed a cytoprotective effect on LPS-induced inflammation in HT-29 cells. We assessed the behavior of LPS-induced cognitive deficit mice using Y-maze, passive avoidance and Morris water maze tests and found that administration of AASWE significantly improved learning and memory function. The AASWE group showed antioxidant activity through downregulation of malondialdehyde levels and upregulation of superoxide dismutase levels in brain tissue. In addition, the AASWE group exhibited activation of the cholinergic system with decreased acetylcholinesterase activity in brain tissue. Furthermore, AASWE effectively downregulated inflammatory mediators such as phosphorylated-JNK, phosphorylated-NF-κB, TNF-α and interleukin-6. The major bioactive compounds of AASWE were identified as quercetin-3-O-arabinopyranosyl(1→2)-rhamnopyranosyl(1→6)-glucopyranose, quercetin-3-O-apiosyl(1 → 2)-galactoside, rutin, and 3-caffeoylquinic acid. Based on these results, we suggest that AASWE not only increases the growth of beneficial bacteria in the intestines, but also shows an ameliorating effect on LPS-induced cognitive impairment.

Amelioration of DSS-Induced Acute Colitis in Mice by Recombinant Monomeric Human Interleukin-22

  • Suhyun Kim;Eun-Hye Hong;Cheol-Ki Lee;Yiseul Ryu;Hyunjin Jeong;Seungnyeong Heo;Joong-Jae Lee;Hyun-Jeong Ko
    • IMMUNE NETWORK
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    • v.22 no.3
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    • pp.26.1-26.18
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    • 2022
  • IL-22, a pleiotropic cytokine, is known to have a profound effect on the regeneration of damaged intestinal barriers. The tissue-protective properties of IL-22 are expected to be potentially exploited in the attenuation and treatment of colitis. However, because of the disease-promoting role of IL-22 in chronic inflammation, a comprehensive evaluation is required to translate IL-22 into the clinical domain. Here, we present the effective production of soluble human IL-22 in bacteria to prove whether recombinant IL-22 has the ability to ameliorate colitis and inflammation. IL-22 was expressed in the form of a biologically active monomer and non-functional oligomers. Monomeric IL-22 (mIL-22) was highly purified through a series of 3 separate chromatographic methods and an enzymatic reaction. We reveal that the resulting mIL-22 is correctly folded and is able to phosphorylate STAT3 in HT-29 cells. Subsequently, we demonstrate that mIL-22 enables the attenuation of dextran sodium sulfate-induced acute colitis in mice, as well as the suppression of pro-inflammatory cytokine production. Collectively, our results suggest that the recombinant mIL-22 is suitable to study the biological roles of endogenous IL-22 in immune responses and can be developed as a biological agent associated with inflammatory disorders.

Probiotic Effects of Lactobacillus plantarum Strains Isolated from Kimchi (김치에서 분리한 Lactobacillus plantarum 균주들의 프로바이오틱 효과)

  • Lee, Xue-Mei;Lee, Hyun Ah;Kweon, Meera;Park, Eui-Seong;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.12
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    • pp.1717-1724
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    • 2016
  • Probiotic effects of Lactobacillus plantarum pF1 NITE-P1462 (Lp-pF1), L. plantarum KCCM 11352P (Lp-PNU), L. plantarum CBT LP3 KCTC 10782BP (Lp-CB), and L. plantarum KCTC 3099 (Lp-3099) isolated from kimchi and Lactococcus lactis KFCC 11510P (L-lactis) isolated from Doenjang were studied. Resistance to gastric and bile acid, adhesion to intestines in colon cells, thermal stability, and antioxidative and in vitro anticancer effects in HT-29 cancer cells were evaluated. L. plantarum strains showed improved tolerance of gastric and bile acids than L-lactis. Lp-pF1 had better adhesion ability in the intestine than Lp-PNU, Lp-3099, and L-lactis. Lp-pF1 also showed better heat resistance at $50^{\circ}C$, $70^{\circ}C$, and $80^{\circ}C$ than Lp-CB, Lp-3099, and L-lactis. In addition, Lp-pF1 exhibited greater antioxidant activity by scavenging DPPH radicals or hydroxyl radicals and anticancer effects in MTT assay than others. Taken together, these results suggest that L. plantarum isolated from kimchi showed higher probiotic activities with antioxidant and anticancer properties than Lac. lactis isolated from Doenjang. Lp-pF1 revealed the best probiotic activities among L. plantarum and could be used as a promising potential probiotics.

Immuno-stimulating and Antitumor Effects of Crude Polysaccharides Extracted from Fruiting Body of Grifola frondosa (잎새버섯(Grifola frondosa)의 자실체에서 추출한 조다당류의 면역증강 및 항암효과)

  • Kim, Jeong-Hwa;Cha, Youn-Jeong;Shim, Mi-Ja;Lee, Min-Woong;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.39 no.1
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    • pp.68-77
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    • 2011
  • 80% methanol and 0.9% neutral saline soluble and hot water substances (hereinafter referred to Fr. NaCl, Fr. HW and Fr. MeOH, respectively) were extracted from fruiting bodies of Grifola frondosa. In vitro cytotoxicity tests, crude polysaccharides were not cytotoxic against cancer cell lines such as Sarcoma 180 and RAW 264.7 at the concentration of 10~2000 ${\mu}g/ml$, but crude polysaccharides from Fr. NaCl was slightly toxic to HT-29 and NIH3T3 at the concentration of 2000 ${\mu}g/ml$. Intraperitoneal injection with crude polysaccharides exhibited life prolongation effect of 25.0~52.9% in mice previously inoculated with Sarcoma 180. Fr. HW increased the numbers of spleen cells by 1.3 fold at the concentration of 200 ${\mu}g/ml$ compared with control. Fr. NaCl improved the immuno-stimulating activity of B lymphocyte by increasing the alkaline phosphatase activity by 1.5 fold compared with control at the concentration of 200 ${\mu}g/ml$. 10~14 ${\mu}M$ of nitric acid were generated when Fr. NaCl was added to RAW 264.7 at the concentration of 50~500 ${\mu}g/ml$, while the control group produced 4.3 ${\mu}M$ of nitric oxide. The Fr. NaCl, Fr. HW and Fr. MeOH increased the production of TNF-${\alpha}$, IL-$1{\beta}$, Il-2 and IL-6 by more than 1.4 times compared with the control group. The Fr. of MeOH increased the numbers of peritoneal exudate cells and circulating leukocytes by 3.0 and 2.0 folds compared with the control at the concentration of 50 ${\mu}g/ml$, respectively. Therefore, the crude polysaccharides extracted from fruiting bodies of Grifola frondosa could improve antitumor activity of mice.

Antioxidant and Anticancer Activities of Water Extracts from Ceramium kondoi (비단풀 물 추출물의 항산화력 및 항암활성)

  • An, Deok-Ho;Cho, Seug-Ja;Jung, Eun-Sil;Lee, Hyun-Jin;Hwang, Ji-Hwan;Park, Eun-Ju;Park, Hae-Ryong;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.10
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    • pp.1304-1308
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    • 2006
  • The present study describes the preliminary evaluation of the antioxidant activity and the cytotoxic effect of Ceramium kondoi. The antioxidant activities and cytotoxic effect of the water extracts were evaluated by total phenolic contents (TPC), DPPH radical scavenging activity (RSA), reducing power (RP), comet assay, and MTT reduction assay. TPC, DPPH RSA, and RP of the extract at the concentration of $1,000{\mu}g/mL$ was $659.2{\mu}M$, 86.0%, and 1.084, respectively, and those were concentration dependent. The $200{\mu}M\;H_2O_2-induced$DNA damage was inhibited by C. kondoi water extract in a dose dependent manner in human leukocytes. The inhibition was by 62.3, 39.8, 24.8% and 16.4% at the concentration of 5, 10, $25{\mu}g/mL$ and $50{\mu}g/mL$, respectively. Cytotoxic activity on HT-29 cells and MCF-7 cells of the C. kondoi water extract at the concentration of $10{\mu}g/mL$ was 49% and 60%, respectively. These results strongly support the possibility of C. kondoi as a source of natural functional materials.