• Title/Summary/Keyword: $A{\beta}$ cell model

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Inhibitory effect of Anglicae dahuricae radix on mast cell mediated inflammatory responses

  • Park, Jong-Ha;Choi, In-Young;Na, Ho-Jeong;Jeong, Hyun-Ja;Ko, Seong-Guem;Lee, Byung-Hee;Song, Yong-Sun;Kim, Hyung-Min;Hong, Seung-Heon;Um, Jae-Young
    • Advances in Traditional Medicine
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    • v.5 no.3
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    • pp.188-194
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    • 2005
  • The Angelicae dahuricae radix (ADR) has been used a traditional medicine to treat acne, erythema, headache, toothache, sinusitis, colds, and flu in Korea, Japan and China. Here, we report the effect of ADR on compound 48/80-induced ear-swelling and the phorbol myristate acetate (PMA) plus calcium ionophore A23187-induced inflammatory cytokine secretion in the human mast cell line, HMC-1. ADR dose-dependently inhibited the ear-swelling response induced by intradermal injection of compound 48/80, In vitro model, PMA plus A23187 significantly increased interleukin $(IL)-1{\beta}$, IL-8, granulocyte macrophage colony stimulating factor (GM-CSF), and tumor necrosis factor $(TNF)-{\alpha}$ secretion compared with media control. We also show that the increased cytokines $IL-1{\beta}$, IL-8, GM-CSF, and $TNF-{\alpha}$ level was significantly inhibited by treatment of ADR. In addition, ADR partially blocked PMA plus A23187-induced extracelluar signal-regulated kinases phosphorylation. These results suggest that ADR might explain its beneficial effect in the treatment of mast cell-mediated inflammatory diseases.

AMPK γ is Required for Maintaining Epithelial Cell Structure and Polarity (AMPK γ 유전자의 표피세포극성 유지기능 규명)

  • Koh, Hyong-Jong
    • Journal of Life Science
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    • v.21 no.5
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    • pp.621-626
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    • 2011
  • AMP-activated protein kinase (AMPK), a heterotrimeric complex comprising a catalytic ${\alpha}$ subunit and regulatory ${\beta}$ and ${\gamma}$ subunits, has been primarily studied as a major metabolic regulator in various organisms, but recent genetic studies discover its novel physiological functions. The first animal model with no functional AMPK ${\gamma}$ subunit gene was generated by using Drosophila genetics. AMPK ${\gamma}$ flies demonstrated lethality with severe defects in cuticle formation. Further histological analysis found that deletion of AMPK ${\gamma}$ causes severe defects in cell polarity in embryo epithelia. The phosphorylation of nonmuscle myosin regulatory light chain (MRLC), a critical regulator of epithelial cell polarity, was also diminished in AMPK ${\gamma}$ embryo epithelia. These defects in AMPK ${\gamma}$ mutant epithelia were successfully restored by over-expression of AMPK ${\gamma}$. Collectively, these results suggested that AMPK ${\gamma}$ is a critical cell polarity regulator in metazoan development.

Inhibitory effect of Alzheimer's in APP-induced Neuro 2A cells by JangWonHwanGagambang(JWHG) water extract (장원환가감방 수추출물(水抽出物)이 아밀로이드 전구단백질으로 유도된 생쥐의 신경아세포주에서의 항치매 효과)

  • Kim, Sang-Tae;Lee, Jong-Hwa;Kim, Tae-Hyeon;Son, Hyeong-Jin;Han, Pyeong-Leem;Kang, Hyung-Won
    • Journal of Oriental Neuropsychiatry
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    • v.18 no.2
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    • pp.13-24
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    • 2007
  • Objective : Recent studies indicate that the deposition of ${\beta}-amyloid$ ($A{\beta}$) is related in the pathogenesis of Alzheimer's disease (AD), but the underlying mechanism is still not clear. Method : To investigate the potential cellular functions of APP and water extract of the JangwonHwangagambang (JWHG), we use as in vitro model, neuro 2A cells were treated with either JWHG or its oriental medicines, and the effect in APP expression was determined by MTT and LDH assay. JWHG have been shown to be neuroprotective in different model systems. We asked whether JWHG treatment would influence cell survival and AD-like pathology in APP-induced neuronal cells. Result : JWHG and water extracts of some oriental medicine has attenuated high cell death in vitro. JWHG-treated cells increased percentage of cell survival more longly than controls. JWHG had significantly increas neurite outgrowth in the as compared to control cells. Conclusion : These results suggest that JWHG prevent APP-induced neurotoxicity through attenuating oxidative stress, and may be useful as potential therapeutic agents for AD.

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Structure of the Starch-Binding Domain of Bacillus cereus $\beta-Amylase$

  • Yoon, Hye-Jin;Akira, Hirata;Motoyasu, Adachi;Atsushi, Sekine;Shigeru, Utsumi;Bunzo, Mikami
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.619-623
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    • 1999
  • The C-terminal starch-binding domain of Bacillus cereus $\beta$-amylase expressed in Escherichia coli was purified and crystallized using the vapor diffusion method. The crystals obtained belong to a space group of $P3_2$ 21 with cell dimensions, a=b=60.20${\AA},\; c=64.92{\AA},\; and \; \gamma = 120^{\circ}$ The structure was determined by the molecular replacement method and refined at 1.95 ${\AA}$, with R-factors of 0.181. The final model of the starch-binding domain comprised 99 amino acid residues and 108 water molecules. The starch-binding domain had a secondary structure of two 4-stranded antiparallel p-sheets similar to domain E of cyclodextrin glucanotransferase and the C-terminal starch-binding domain of glucoamylase. A comparison of the structures of these starch-binding domains revealed that the separated starch-binding domain of Bacillus cereus $\beta-Amylase$had only one starch-binding site (site 1) in contrast to two sites (site 1 and site 2) reported in the domains of cyclodextrin glucanotransferase and glucoamylase.

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Mammary alveolar cell as in vitro evaluation system for casein gene expression involved in glucose level

  • Heo, Young Tae;Ha, Woo Tae;Lee, Ran;Lee, Won-Young;Jeong, Ha Yeon;Hwang, Kyu Chan;Song, Hyuk
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.6
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    • pp.878-885
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    • 2017
  • Objective: Glucose is an essential fuel in the energy metabolism and synthesis pathways of all mammalian cells. In lactating animals, glucose is the major precursor for lactose and is a substrate for the synthesis of milk proteins and fat in mammary secretory (alveolar) epithelial cells. However, clear utilization of glucose in mammary cells during lactogenesis is still unknown, due to the lack of in vitro analyzing models. Therefore, the objective of this study was to test the reliability of the mammary alveolar (MAC-T) cell as an in vitro study model for glucose metabolism and lactating system. Methods: Undifferentiated MAC-T cells were cultured in three types of Dulbecco's modified Eagle's medium with varying levels of glucose (no-glucose: 0 g/L, low-glucose: 1 g/L, and high-glucose: 4.5 g/L) for 8 d, after which differentiation to casein secretion was induced. Cell proliferation and expression levels of apoptotic genes, Insulin like growth factor-1 (IGF1) receptor, oxytocin receptor, ${\alpha}S1$, ${\alpha}S2$, and ${\beta}$ casein genes were analyzed at 1, 2, 4, and 8 d after differentiation. Results: The proliferation of MAC-T cells with high-glucose treatment was seen to be significantly higher. Expression of apoptotic genes was not affected in any group. However, expression levels of the mammary development related gene (IGF1 receptor) and lactation related gene (oxytocin receptor) were significantly higher in the low-glucose group. Expressions of ${\alpha}S1-casein$, ${\alpha}S2-casein$, and ${\beta}-casein$ were also higher in the low-glucose treated group as compared to that in the no-glucose and high-glucose groups. Conclusion: The results demonstrated that although a high-glucose environment increases cell proliferation in MAC-T cells, a low-glucose treatment to MAC-T cells induces higher expression of casein genes. Our results suggest that the MAC-T cells may be used as an in vitro model to analyze mammary cell development and lactation connected with precise biological effects.

Anti-inflammatory effect of the water fraction from hawthorn fruit on LPS-stimulated RAW 264.7 cells

  • Li, Chunmei;Wang, Myeong-Hyeon
    • Nutrition Research and Practice
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    • v.5 no.2
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    • pp.101-106
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    • 2011
  • The hawthorn fruit (Crataegus pinnatifida Bunge var. typica Schneider) is used as a traditional medicine in Korea. The objective of this study was to understand the mechanisms of the anti-inflammatory effects of the water fractionated portion of hawthorn fruit on a lipopolysaccharide (LPS)-stimulated RAW 264.7 cellular model. The level of nitric oxide (NO) production in the water fraction and LPS-treated RAW 264.7 cells were determined with an ELISA. The cytotoxicity of the water fraction and LPS was measured with an MTT assay. Expression of nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor (TNF)-${\alpha}$, interleukin 6 (IL-6), and interleukin $1{\beta}$ (IL-$1{\beta}$) mRNA were analyzed with a reverse transcription polymerase chain reaction (RT-PCR). The water fraction of hawthorn fruit was determined to be safe and significantly inhibited NO production in LPS-stimulated RAW 264.7 cells and suppressed COX-2, (TNF)-${\alpha}$, IL-$1{\beta}$, and IL-6 expression. The observed anti-inflammatory effects of the water fraction of hawthorn fruit might be attributed to the down-regulation of COX-2, (TNF)-${\alpha}$, IL-$1{\beta}$, and IL-6 expression in LPS-stimulated RAW 264.7 cells.

Enhancement of Immune Activities of Canavalia gladiata & Arctium lappa complexes in immobilization stress mouse model. (부동화 스트레스 유도 마우스 모델에서 도두(刀豆), 우방근(牛蒡根) 복합물의 면역증진 작용)

  • Lee, Ji-eun;Roh, Seong-Soo;Kim, Han-Young;Kim, Kun-hoae;Kim, Seung-Hyung
    • The Korea Journal of Herbology
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    • v.32 no.1
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    • pp.1-13
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    • 2017
  • Objective : Soybeans of Canavalia gladiata(CG) and root of Arctium lappa(AL) have been reported to have anti-inflammatory, antioxidant effect. However, the immunoregulatory mechanisms of its combinational prescription remain a matter of considerable debate. In the current study, we investigated whether CG and AL and its combinational prescription(CG+AL) regulate immune system using chronic immobilization-stress mouse model. Methods : C57BL/6J mice fixed for 2 hours into immobilization tube after CG, AL, CG+AL oral administration after 2 hours daily for 21 days. After every experiment has ended the C57BL/6J mice were sacrificed on 22 days. The production of Serotonin and Cortisol, lgA were observed by ELISA method, The proportion of immune cells such as T/B cell and macrophage, NK cell were measured by FACS. Then, Real-time PCR was performed to measure the mRNA expression of Inflammatory cytokines(IL-1beta, IL-6, TNF-a) and T cell activation cytokines(IL-2, IL-10, IFN-gamma, IL-12p35 / p40). Result : When chronic immobilization-stress mouse model were treated with CG+AL(1:4), the expression of mRNA were significantly decreased at the Inflammatory cytokines(IL-1beta, IL-6, TNF-a). While, the levels of mRNA were significantly increased at immune T cell activation cytokines. Additionally, CG+AL(1:4) combinational prescription group enhanced immune cells such as T/B cell and macrophage, NK cell. Furthermore, the Immuno-fluorescence result of brain tissue can confirm that CG+AL(1:4) group significantly increased the BDNF expression. Conclusion : These result suggest that CG+AL(1:4) combinational prescription has Immune System enhancement via stress-mediated immunocyte.

Effect of ${\beta}$-glucan Originated from Aureobasidium on the Dermal Wound Repair in Vitro Model (생체외 모델에서 아우레오바지디움 유래 베타 글루칸이 피부창상 치유에 미치는 영향)

  • Lee, Jung-Woo;Kwon, Young-Sam;Jang, Kwang-Ho
    • Journal of Veterinary Clinics
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    • v.30 no.6
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    • pp.403-408
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    • 2013
  • The objective of the present study is to detect the effect of ${\beta}$-glucan originated from Aureobasidium on the proliferation and collagen production in human dermal fibroblast cells with wound repopulation in vitro. The proliferative effects were assessed using a MTT assay as well as cell counts at 24 and 48 hr after treatment. Hydroxyproline was measured as an index of procollagen production with reverse-phase high pressure liquid chromatography. Oncostatin M was used as a reference agent. In glucagon treated group, dose-dependent and significant increase of optical density or fibroblast cell numbers was demonstrated, when compared with those of control from 0.1 mg/ml concentration. In addition, the numbers of cells which had migrated into the wound defects were more significantly and dose-dependently increased than those of non-treated control. However, no meaningful effects on the procollagen production were observed.

Anti-bacterial effects of enzymatically-isolated sialic acid from glycomacropeptide in a Helicobacter pylori-infected murine model

  • Noh, Hye-Ji;Koh, Hong Bum;Kim, Hee-Kyoung;Cho, Hyang Hyun;Lee, Jeongmin
    • Nutrition Research and Practice
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    • v.11 no.1
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    • pp.11-16
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    • 2017
  • BACKGROUND/OBJECTIVES: Helicobacter pylori (H. pylori) colonization of the stomach mucosa and duodenum is the major cause of acute and chronic gastroduodenal pathology in humans. Efforts to find effective anti-bacterial strategies against H. pylori for the non-antibiotic control of H. pylori infection are urgently required. In this study, we used whey to prepare glycomacropeptide (GMP), from which sialic acid (G-SA) was enzymatically isolated. We investigated the anti-bacterial effects of G-SA against H. pylori in vitro and in an H. pylori-infected murine model. MATERIALS/METHODS: The anti-bacterial activity of G-SA was measured in vitro using the macrodilution method, and interleukin-8 (IL-8) production was measured in H. pylori and AGS cell co-cultures by ELISA. For in vivo study, G-SA 5 g/kg body weight (bw)/day and H. pylori were administered to mice three times over one week. After one week, G-SA 5 g/kg bw/day alone was administered every day for one week. Tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), IL-$1{\beta}$, IL-6, and IL-10 levels were measured by ELISA to determine the anti-inflammatory effects of G-SA. In addition, real-time PCR was performed to measure the genetic expression of cytotoxin-associated gene A (cagA). RESULTS: G-SA inhibited the growth of H. pylori and suppressed IL-8 production in H. pylori and in AGS cell co-cultures in vitro. In the in vivo assay, administration of G-SA reduced levels of IL-$1{\beta}$ and IL-6 pro-inflammatory cytokines whereas IL-10 level increased. Also, G-SA suppressed the expression of cagA in the stomach of H. pylori-infected mice. CONCLUSION: G-SA possesses anti-H. pylori activity as well as an anti-H. pylori-induced gastric inflammatory effect in an experimental H. pylori-infected murine model. G-SA has potential as an alternative to antibiotics for the prevention of H. pylori infection and H. pylori-induced gastric disease prevention.

The Effect of Jeongshin-tang on Interleukin-1 $\beta$ and $\beta$-Amyloid-Induced Cytokine Production in Human Brain Astrocytes

  • Kim Bo Kyung;Shin Soon Shik;Kang Seon Tae
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.1
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    • pp.254-259
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    • 2004
  • Jeongshin-tang (JST) is a Korean herbal prescription, which has been successfully applied for the various neuronal diseases. However, it's effect remains unknown in experimental models. To investigate the biological effect of JST in Alzheimer's disease (AD) in vitro model, we analized the production of interleukin (IL)-6 and IL-8, and expression of cyclooxygenase (COX)-2 in IL-1β plus β-amyloid [25-35] fragment (A)-stimulated human astrocytoma cell line U373MG. JST alone had no effect on the cell viability. The production of IL-6 and IL-8 was significantly inhibited by pretreatment with JST (1mg/㎖) on IL-1β plus A-stimulated U373MG cells. Maximal inhibition rate of IL-6 and IL-8 production by JST was about 41.22% (P<0.01) and 34.45% (P<0.05), respectively. The expression level of COX-2 protein was up-regulated by IL-1β plus A but the increased level of COX-2 was inhibited by pretreatment with JST (1 mg/㎖). These data indicate that JST has a regulatory effect on cytokine production and COX-2 expression, which might explain it's beneficial effect in the treatment of AD.