• Title/Summary/Keyword: Enzyme inhibitor

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Characteristics and Physiological Functionalities of Unrecorded Yeasts from Wild Flowers of Seonyudo in Jeollabuk-do, Korea (전라북도 선유도일대 야생화에서 분리한 국내 미기록 효모들의 특성 및 생리기능성)

  • Hyun, Se-Hee;Han, Sang-Min;Lee, Jong-Soo
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.402-406
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    • 2014
  • Six kinds of newly recorded yeasts such as Rhodosporidium diobovatum SY4-2, Cryptococcus bestiolae SY7-1, Kazachstania unispora SY14-1, Kazachstania servazzii SY14-3, Pichia holstii SY20-2 and Cryptococcus tephrensis SY26-1 were screened from sixty one yeasts derived from wild flowers found in Seonyudo, Gogunsanyeoldo, Jeollabuk-do, Korea. All of them grew in 50% glucose-containing yeast extract-peptone-dextrose (YPD) broth and Pichia holstii SY20-2 was also halophile, growing in 20% NaCl-containing YPD broth. All of them, except Cryptococcus tephrensis SY26-1, were assimilated to glucose. Cell-free extract from Kazachstania servazzii SY14-3 showed the highest 98.6% of ${\alpha}$-glucosidase inhibitory activity and maximal production of the ${\alpha}$-glucosidase inhibitor was obtained with 24h incubation at $30^{\circ}C$. The antihypertensive angiotensin I-converting enzyme inhibitory activity of the unrecorded yeasts were showed 58.6-80.4% in their supernatants.

Studies on the Anti-inflammatory and Anti-apoptotic Effect of Catalposide Isolated from Catalpa ovata (개오동나무에서 추출(抽出)한 Catalposide의 항염(抗炎) 및 세포고사(細胞枯死) 억제효과(抑制效果)에 관(關)한 연구(硏究))

  • Oh, Cheon-Sik;Hwang, Sang-Wook;Kim, Yong-Woo;Song, Dal-Soo;Chae, Young-Seok;Jeong, Jong-Gil;Song, Ho-Joon;Shin, Min-Kyo
    • The Korea Journal of Herbology
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    • v.20 no.3
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    • pp.29-41
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    • 2005
  • Objectives : The use of natural products with therapeutic properties is as ancient as human civilisation and, for a long time, mineral, plant and animal products were the main sources of drugs. Catalposide, the major iridoid glycoside isolated from the stem bark of Catalpa ovata G. Don (Bignoniceae) has been shown to possess anti-microbial and anti-tumoral properties. Heme oxygenase-1 (HO-1) is a stress response protein and is known to play a protective role against the oxidative injury. In this study, we examined whether catalposide could protect Neuro 2A cells, a kind of neuronal cell lines, from oxidative damage through the induction of HO-1 protein expression and HO activity. We also examined the effects of catalposide on the productions of tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ and nitric oxide (NO) on RAW 264.7 macrophages activated with the endotoxin lipopolysaccharide. Methods : HO-1 expression in Neuro 2A cells was measured by Western blotting analysis. NO and $TNF--{\alpha}$ produced by RAW 264.7 macrophage were measured by Griess reagent and enzyme-linked immunosorbent assay, respectively. Results : The treatment of the cells with catalposide resulted in dose- and time-dependent up-regulations of both HO-1 protein expression and HO activity. Catalposide protected the cells from hydrogen peroxide-induced cell death. The protective effect of catalposide on hydrogen peroxide-induced cell death was abrogated by zinc protoporphyrin IX, a HO inhibitor. Additional experiments revealed the involvement of CO in the cytoprotective effect of catalposide-induced HO-1. In addition, catalposide inhibited the productions of $TNF--{\alpha}$ and NO with significant decreases in mRNA levels of $TNF--{\alpha}$ and inducible NO synthase. Conclusions : Our results indicate that catalposide is a potent inducer of HO-1 and HO-1 induction is responsible for the catalposide-mediated cytoprotection against oxidative damage and that catalposide may have therapeutic potential in the control of inflammatory disorders.

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Vitamin D Inhibits Expression and Activity of Matrix Metalloproteinase in Human Lung Fibroblasts (HFL-1) Cells

  • Kim, Seo Hwa;Baek, Moon Seong;Yoon, Dong Sik;Park, Jong Seol;Yoon, Byoung Wook;Oh, Byoung Su;Park, Jinkyeong;Kim, Hui Jung
    • Tuberculosis and Respiratory Diseases
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    • v.77 no.2
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    • pp.73-80
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    • 2014
  • Background: Low levels of serum vitamin D is associated with several lung diseases. The production and activation of matrix metalloproteinases (MMPs) may play an important role in the pathogenesis of emphysema. The aim of the current study therefore is to investigate if vitamin D modulates the expression and activation of MMP-2 and MMP-9 in human lung fibroblasts (HFL-1) cells. Methods: HFL-1 cells were cast into three-dimensional collagen gels and stimulated with or without interleukin-$1{\beta}$ (IL-$1{\beta}$) in the presence or absence of 100 nM 25-hydroxyvitamin D (25(OH)D) or 1,25-dihydroxyvitamin D ($1,25(OH)_2D$) for 48 hours. Trypsin was then added into the culture medium in order to activate MMPs. To investigate the activity of MMP-2 and MMP-9, gelatin zymography was performed. The expression of the tissue inhibitor of metalloproteinase (TIMP-1, TIMP-2) was measured by enzyme-linked immunosorbent assay. Expression of MMP-9 mRNA and TIMP-1, TIMP-2 mRNA was quantified by real time reverse transcription polymerase chain reaction. Results: IL-$1{\beta}$ significantly stimulated MMP-9 production and mRNA expression. Trypsin converted latent MMP-2 and MMP-9 into their active forms of MMP-2 (66 kDa) and MMP-9 (82 kDa) within 24 hours. This conversion was significantly inhibited by 25(OH)D (100 nM) and $1,25(OH)_2D$ (100 nM). The expression of MMP-9 mRNA was also significantly inhibited by 25(OH)D and $1,25(OH)_2D$. Conclusion: Vitamin D, 25(OH)D, and $1,25(OH)_2D$ play a role in regulating human lung fibroblast functions in wound repair and tissue remodeling through not only inhibiting IL-$1{\beta}$ stimulated MMP-9 production and conversion to its active form but also inhibiting IL-$1{\beta}$ inhibition on TIMP-1 and TIMP-2 production.

Adipose-Derived Stem Cells Suppress Inflammation Induced by IL-1β through Down-Regulation of P2X7R Mediated by miR-373 in Chondrocytes of Osteoarthritis

  • Jin, Rilong;Shen, Miaoda;Yu, Liedao;Wang, Xuanwei;Lin, Xiangjin
    • Molecules and Cells
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    • v.40 no.3
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    • pp.222-229
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    • 2017
  • Adipose-derived stem cells (ADSCs) were previously considered to have an anti-inflammatory effect, and Interleukin-$1{\beta}$ ($IL-1{\beta}$) was found to be a pro-inflammatory factor in chondrocytes, but the mechanism underlying ADSCs and $IL-1{\beta}$ is unclear. In this study, we investigate whether P2X7 receptor (P2X7R) signalling, regulated by microRNA 373 (miR-373), was involved in the ADSCs and $IL-1{\beta}$ mediated inflammation in osteoarthritis (OA). Chondrocytes were collected from 20 OA patients and 20 control participants, and ADSCs were collected from patients who had undergone abdominal surgery. The typical surface molecules of ASDCs were detected by flow cytometry. The level of nitric oxide (NO) was determined by Griess reagent. Concentrations of prostaglandin E2 (PGE2), interleukin 6 (IL-6), matrix metallopeptidase 3 (MMP-3) were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of IL-6, MMP-3, miR-373 and P2X7R were determined by real-time polymerase chain reaction (PCR), and Western blot was used to detect the protein expression of P2X7R. The typical potential characters of ADSCs were verified. In chondrocytes or OA tissues, the miR-373 expression level was decreased, but the P2X7R expression was increased. $IL-1{\beta}$ stimulation increased the level of inflammatory factors in OA chondrocytes, and ADSCs co-cultured with $IL-1{\beta}$-stimulated chondrocytes decreased the inflammation. OA chondrocytes transfected with the miR-373 inhibitor increased the inflammation level. The miR-373 mimic suppressed the inflammation by targeting P2X7R and regulated its expression, while its effect was reversed by overexpression of P2X7R. $IL-1{\beta}$ induced inflammation in OA chondrocytes, while ADSCs seemed to inhibit the expression of P2X7R that was regulated by miR-373 and involved in the anti-inflammatory process in OA.

Acetylcholinesterase Inhibitory Effect of Green Tea Extracts According to Storage Condition (저장 조건에 따른 녹차 추출물의 acetylcholinesterase(AChE) 저해 효과)

  • Kwak, Ji Hyun;Jeong, Chang-Ho;Kim, Ji Hye;Choi, Gwi Nam;Shin, Young-Hee;Lee, Seung-Cheol;Cho, Sung Hwan;Choi, Sung-Gil;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.41 no.4
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    • pp.435-440
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    • 2009
  • Acetylcholinesterase (AChE) inhibitors, which enhance cholinergic transmission by reducing the enzymatic degradation of acetylcholine, are the only source of the compound that is currently approved for the treatment of Alzheimer's disease (AD). In these experiments, the concentration samples of green tea extracts were 100, 500, and 1,000 ${\mu}g$/mL. Among them, the highest concentration (1,000 ${\mu}g$/mL) of fresh green tea extracts showed the most potent inhibitory effect on AChE by reducing more than 40% of enzyme activity, and in a concentration-dependent pattern. In addition, we examined the effect of other storing conditions on AChE inhibition. In order to maintain storage for 3 months, the materials were held at the certain storing conditions of temperature (room temperature, 4 and $-20^{\circ}C$) and for water activity (0.81, 0.69, and 0.23). In these storing conditions, the difference in temperature did not contribute to the AChE inhibitory effect. Our presented data showed that the AChE inhibitory effect was affected by the concentration of green tea extract and by water activity (0.81). These results suggest that green tea may serve as a potential dietary source of AChE inhibitor.

Pre-and Post-ishemic Changes of the Constituent Enzymes in Isolated Rabbit's Myocardium (허혈전후 적출 가토 심근내의 구성 효소의 변화)

  • 천수봉;전도환;이재성;김송명
    • Journal of Chest Surgery
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    • v.33 no.2
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    • pp.117-124
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    • 2000
  • Background: Nucleoside transport inhibitor(NTI) Keeps AMP, ADP, ATP levels high in myocytes by inhibiting adenosine cataboilsm so that it may preserve the myocardial contractability during ischemia In this study we investigated the effects of cyclic AMP phosphodiesterase inhibor(C-AMP PDSI) and S-P-nitrobenzyl-6 -thioniosine(NBT; a sort of NIT) on myocadial preservation and changes of constituent enzyme. Material and method: Twenty-six isolated rabbit hearts were perfused with Krebs-Henseleit buffer solution for 20 minutes arrested for 20 minutes and ten reperfused for 30 minutes. The following four groups were prepared and hemodynamic changes coronary effluent lactate dehydrogenase (LDH) a-hydroxybutylic accid(a-HBD) levels and myocardial LDH creatine kinase-MB (CK-MB) adenosine deaminase(ADA) a-HBD levels and myocardial LDH creatine kinase-MB (CK-MB) adenosine deaminase(ADA) a-HBD levels were analysed before and after cardiac arest ; Group I(control) ; the heart was only perfused with K-H ; Group II ; the heart was perfused with K-H including C-AMP PDSI(Amrinone 25mg/L); Group III ; the heart was perfused with K-H including NBT(4.19mg/L) ; Group IV ; the heart was perfused with K-H including C-AMP PDSI + NBT. Result : Left venticular developed pressure(LVDP) at 10 minutes of the equilibrium was significantly higher in group III(72.1$\pm$5.3 mmHg p<0.01) and group III(72$\pm$5.6 mmHg P<0.025) as compared with group I (40.8$\pm$4.7mmHg) and LVDP at 20 minutes of the reperfusion was significantly higher in group II(74$\pm$5.3mmHg p<0.01) and group III(72$\pm$5.6mmHg p<0.025) as compared with group I (44.2$\pm$4.6mmHg). Percentage recovery of LVDP at the reperfusion was the highest in group II(123.3%) Percentage recovery of coronary flow at the equilibrium reperfusion were higher in group II(310%, 270%) group III(230%, 290%) group IV(310%, 280%) as compared with group I (100%) respectively. Myocadial LDH level was significant lower in group IV(33495$\pm$1802 IU/gm p<0.04) as compared with group I(48767$\pm$1421 IU/gm) Myocadial CK-MB level was significant higher in group II(74820$\pm$1421 IU/gm) compared with group I (45450$\pm$1737 IU/gm) Myocadial ADA level was significant higher group IV(1215$\pm$8 IU/gm p<0.05) compared with group I(125$\pm$15 IU/gm) but there was no significant difference between group I and group II ,III, IV in changes of coronary effluent LDH, a-HBD levels. Conclusion: C-AMP PDSI solely appears to have a better effect on myocardial preservation after ischemia than NBT but with no synergistic effect and it could keep CK-MB leve high in myocardial tissues.

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Study on brewing of sweet potato starch (고구마 전분질원료를 이용한 주류제조에 관한 연구)

  • 정기택;유대식
    • Korean Journal of Microbiology
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    • v.9 no.3
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    • pp.103-120
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    • 1971
  • We have been studied on brewing sweet starch. We obtained the results as follows ; 1) 5 strains, T-T-2, T-T-4, T-K-2, T-T-18, T-T-1, were the most available in view of fermentative power by capacity of $CO_2$. 2) 5 strains, T-T-4, T-T-2, T-T-1, T-T-3, T-K-2, produced capacity of alcohol more than 5.78%. 3) 6 strains, T-T-2, T-K-2, T-T-4, T-S-2, T-I-3, T-I-1, are available not only taste and flavour, but productive power of alcohol in sweet potato starch. 4) The form of 6 strains are long oval and round and most of them are similar to the other yeast in size. 5) In giant colony the color was cream color and cream buff, and T-K-2 was formed by $15{\times}12mm$ on diameter and by 3.5mm on high. 6) Optimum temperature of most of all strains is 25~ $300^{\circ}C$but T-K-4 is 28-30.deg.C. 7) Optimum pH is 3.4-4.6. 8) T-S-2 was died off at 65.deg.C, the other strains died $60^{\circ}C$. 9( Making Bun-kok with non-heated wheat bran .alpha.-amylase was more increased by 4.5-13.5 mg of glucose in reaction solution and .betha.-amylase more 1.6-3.4ml of N/10-$KMnO_4$ Solution than Bun-kok with heated wheat bran. 10) It seems that mycellium grows better than original in substance containing 0.4 ~ 1.2% of HCl. 11) Making Bun-kok to add 0.8% HCl, .alpha.-amylase was increased 9.93-20.7mg of glucose and .betha.-amylase ws increased 2.6~4.3ml of N/10-$KMnO_4$ solution to reaction solution. 12) 1.2%-HCl, or higher concentration, acts as inhibitor, in the meanwhile the concentration between 0.4~0.8% of HCl acts as activator. 13) We must make Bun-kok for 42 hours, at 28~$30^{\circ}C$) After we made Bun-kok using S-O-II and R-J-I one by one, Bun-kok which mix each other in equal quantity is increased more than original on enzyme acrivity. 15) Oxidation is the best way of refining sweet potato starch in N/10-phosphate buffer solution (pH 7.5). 16) When we prepared sweet potato starch, first pH was 3.0.

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Transglutaminase-2 Is Involved in All-Trans Retinoic Acid-Induced Invasion and Matrix Metalloproteinases Expression of SH-SY5Y Neuroblastoma Cells via NF-κB Pathway

  • Lee, Hye-Ja;Park, Mi-Kyung;Bae, Hyun-Cheol;Yoon, Hee-Jung;Kim, Soo-Youl;Lee, Chang-Hoon
    • Biomolecules & Therapeutics
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    • v.20 no.3
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    • pp.286-292
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    • 2012
  • All-trans retinoic acid (ATRA) is currently used in adjuvant differentiation-based treatment of residual or relapsed neuroblastoma (NB). It has been reported that short-term ATRA treatment induces migration and invasion of SH-SY5Y via transglutaminase-2 (Tgase-2). However, the detailed mechanism of Tgase-2's involvement in NB cell invasion remains unclear. Therefore we investigated the role of Tgase-2 in invasion of NB cells using SH-SY5Y cells. ATRA dose-dependently induced the invasion of SH-SY5Y cells. Cystamine (CTM), a well known tgase inhibitor suppressed the ATRA-induced invasion of SH-SY5Y cells in a dose-dependent manner. Matrix metalloproteinase -9 (MMP-9) and MMP-2, well known genes involved in invasion of cancer cells were induced in the ATRA-induced invasion of the SH-SH5Y cells. Treatment of CTM suppressed the MMP-9 and MMP-2 enzyme activities in the ATRA-induced invasion of the SH-SY5Y cells. To confirm the involvement of Tgase-2, gene silencing of Tgase-2 was performed in the ATRA-induced invasion of the SH-SH5Y cells. The siRNA of Tgase-2 suppressed the MMP-9 and MMP-2 activity of the SH-SY5Y cells. MMP-2 and MMP-9 are well known target genes of NF-${\kappa}B$. Therefore the relationship of Tgase-2 and NF-${\kappa}B$ in the ATRA-induced invasion of the SH-SY5Y cells was examined using siRNA and CTM. ATRA induced the activation of NF-${\kappa}B$ in the SH-SY5Y cells and CTM suppressed the activation of NF-${\kappa}B$. Gene silencing of Tgase-2 suppressed the MMP expression by ATRA. These results suggested that Tgase-2 might be a new target for controlling the ATRA-induced invasion of NBs.

Effects of (-)-Epigallocatechin-3-gallate on the Release of Pancreatic Enzymes and Expression of Regenerating Genes in Ethanol-injured Murine Pancreatic Primary Acinar Cells (에탄올에 의하여 유도된 마우스 췌장 선포세포의 염증성 손상에서 췌장분비 효소의 활성 및 세포 재생관련 유전자들의 발현에 미치는 EGCG의 영향)

  • Kim, Sung Ok;Choi, Yung Hyun
    • Journal of Life Science
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    • v.23 no.11
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    • pp.1404-1408
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    • 2013
  • (-)-Epigallocatechin-3-gallate (EGCG), a green tea polyphenol, has been shown to have strong antibacterial, antiviral, antioxidant, anti-inflammatory, and chemopreventive effects. However it is unknown whether EGCG can recover alcohol-associated pancreatitis. The aim of this study was to investigate the effects of EGCG on pancreatic enzyme activities and the expressions of pancreatic regenerating related markers, such as adenosine monophosphate-activated protein kinase (AMPK), raf-1 kinase inhibitor protein (RKIP), and Regenerating gene 1 (Reg1), in mice pancreatic primary acinar cells. Our results revealed that activities of ${\alpha}$-amylase and chymotrypsin were significantly increased in the cells treated with ethanol compared to the untreated control cells; however, the increased activities of both enzymes were markedly reduced by pretreatment with EGCG. Phosphorylation of AMPK and total expression of RKIP were decreased in the ethanol-treated primary acinar cells; however, these were both significantly increased in the EGCG-pretreated cells. In addition, when EGCG was treated, expression of Reg1 was markedly increased compared with that of the control or the ethanol-treated primary acinar cells, demonstrating that EGCG can modulate pancreatic regenerating related genes. Therefore, our findings suggest that EGCG may have therapeutic utility in the prevention or treatment of alcohol-associated pancreatitis.

Physiological Effects of Casein-derived Bioactive Peptides (카제인 유래 생리활성 Peptide의 체내 효과)

  • Jung, Ho-Jung;Min, Bock-Ki;Kwak, Hae-Soo
    • Food Science of Animal Resources
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    • v.29 no.6
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    • pp.659-667
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    • 2009
  • Casein is considered to be the main source of protein in milk; therefore, many studies have been conducted to identify casein-derived bioactive peptides and their physiological effects. Casein is inactive within the parent protein but can be liberated by various proteases and enzymatic hydrolysis during microbial fermentation and gastrointestinal digestion. Once absorbed, casein exhibits different bioavailabilities in the body. Specifically, casein-derived peptides function as angiotensin converting enzyme (ACE) inhibitor in the cardiovascular system; thus, they are expected to reduce and prevent hypertension. Additionally, casein-derived peptides behave as opioid-like peptides in the nervous system, which impacts relaxation. These peptides are also expected to modulate various aspects of immune functions. Finally, caseinophosphopeptide (CPP) and glycomacropeptide (GMP) may exhibit a number of nutritional effects such as the absorption of calcium, iron or zinc. Many studies have been conducted to evaluate casein-derived peptides due to their multifunctional properties and the results of these studies have contributed to the development of a wide variety of functional dairy products. The purpose of this paper was to review the generation of bioactive peptides, their absorption and metabolism, and their specific bioactive effects.