• Title/Summary/Keyword: Degradation factor

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Neuroprotective Effects of Cheongnoemyeongsin-hwan against Hydrogen Peroxide-induced DNA Damage and Apoptosis in Human Neuronal-Derived SH-SY5Y Cells (인체 신경세포에서 청뇌명신환(淸腦明神丸)의 산화적 스트레스에 대한 세포보호 효과)

  • Pi, Guk Hyun;Hwang, Won Deuk
    • Herbal Formula Science
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    • v.25 no.1
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    • pp.51-68
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    • 2017
  • Objectives : Oxidative stress due to excessive accumulation of reactive oxygen species (ROS) is one of the risk factors for the development of several chronic diseases, including neurodegenerative diseases. Methods : In the present study, we investigated the protective effects of cheongnoemyeongsin-hwan (CNMSH) against oxidative stress‑induced cellular damage and elucidated the underlying mechanisms in neuronal-derived SH-SY5Y cells. Results : Our results revealed that treatment with CNMSH prior to hydrogen peroxide (H2O2) exposure significantly increased the SH-SY5Y cell viability, indicating that the exposure of the SH-SY5Y cells to CNMSH conferred a protective effect against oxidative stress. CNMSH also effectively attenuated H2O2‑induced comet tail formation, and decreased the phosphorylation levels of the histone ${\gamma}H2AX$, as well as the number of apoptotic bodies and Annexin V‑positive cells. In addition, CNMSH exhibited scavenging activity against intracellular ROS generation and restored the mitochondria membrane potential (MMP) loss that were induced by H2O2, suggesting that CNMSH prevents H2O2‑induced DNA damage and cell apoptosis. Moreover, H2O2 enhanced the cleavage of caspase-3 and degradation of poly (ADP-ribose)-polymerase, a typical substrate protein of activated caspase-3, as well as DNA fragmentation; however, these events were almost totally reversed by pretreatment with CNMSH. Furthermore, CNMSH increased the levels of heme oxygenase-1 (HO-1), which is a potent antioxidant enzyme, associated with the induction of nuclear factor-erythroid 2-related factor 2 (Nrf2). According to our data, CNMSH is able to protect SH-SY5Y cells from H2O2-induced apoptosis throughout blocking cellular damage related to oxidative stress through a mechanism that would affect ROS elimination and activating Nrf2/HO-1 signaling pathway. Conclusions : Therefore, we believed that CNMSH may potentially serve as an agent for the treatment and prevention of neurodegenerative diseases caused by oxidative stress.

Inhibitory Effects of Ssangbohwan on Osteoclast Differentiation and Bone Resorption (쌍보환 추출물의 파골세포 분화 억제와 골 흡수 억제효과)

  • Kim, Seong Joung;Lee, Jeong Ju;Kim, June Hyun;Jo, So Hyun;Park, Min Cheol;Jo, Eun Heui
    • Journal of Acupuncture Research
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    • v.32 no.3
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    • pp.69-81
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    • 2015
  • Purpose : The first purpose of this study is to find out whether the water extract of Rehmanniae Radix Preparat(RRP), Cuscutae Semen(CS) and their combination(Ssangbohwan, SBH) have the effect of suppressing Receptor activator of nuclear factor kappa-B ligand(RANKL)-induced osteoclast differentiation. The second purpose of this study is to find out whether the water extract of RRP, CS and SBH have the effect of inhibiting osteoporosis in an osteoporosis model induced by lipopolysaccharide(LPS). Methods : After promoting differentiation of osteoclasts by treating the RANKL, we observed the effect by the administration of RRP, CS and SBH. In addition, by means of Reverse transcription polymerase chain reaction(RT-PCR), we assayed mRNA expression levels of NFATc1, c-Fos, TRAP and GAPDHS(Glyceraldehyde-3-phosphate dehydrogenase, spermatogeni) from bone marrow macrophages(BMMs). Similarly, the protein expression levels of NFATc1 (Nuclear factor of activated T-cells, cytoplasmic1), C-Fos, MAPKs(Mitogen-activated protein kinases) and ${\beta}$-actin in cell lysates were analyzed by means of Western Blotting. Finally, we determined the anti-osteoporotic effects of RRP, CS and SBH, through the use of Lipopolysaccharide-induced bone-loss mouse. Results : RRP, CS and SBH showed remarkable inhibitive effect on RANKL-treated osteoclast differentiation without cytotoxicity. SBH inhibited the phosphorylation of p38, Jun N-terminal kinases(JNK), and I-${\kappa}B$ and down-regulated the induction of c-Fos and NFATc1 by RANKL. RRP, CS suppressed degradation of I-${\kappa}B$, but it did not affect c-Fos and NFATc1 by RANKL. Lastly, in vivo data showed that RRP and SBH prevented bone erosion by LPS treatment. Conclusions : These results demonstrate SBH can be effective remedy for bone-loss diseases such as osteoporosis.

The anti-allergy and anti-inflammatory effect of Anemarrhenae Rhizoma in vivo and in vitro

  • Kim, Su-Jin;Jeong, Hyun-Ja;Myung, Noh-Yil;Moon, Phil-Dong;Lee, Ju-Young;Yi, Byoung-Jae;Lee, Eun-Hyub;An, Nyeon-Hyung;Park, Seok-Jae;Kim, Min-Cheol;Jun, Suk-Min;Lee, Ji-Hyun;Kim, Hyung-Min;Hong, Seung-Heon;Um, Jae-Young
    • Advances in Traditional Medicine
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    • v.7 no.3
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    • pp.235-243
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    • 2007
  • Anemarrhenae Rhizoma (AR) is used in traditional oriental medicine for various medicinal purposes. However, the exact mechanism that accounts for the anti-allergy and anti-inflammatory effects of the AR is still not fully understood. The aim of The present study is to elucidate whether and how AR modulates the allergic reactions in vivo, and inflammatory reaction in vitro. In this study, we showed that AR significantly decreased compound 48/80-induced systemic anaphylaxis, paw oedema, and histamine release from preparation of rat peritoneal mast cells. Also, AR inhibited the expression of inflammatory cytokine in PMA plus A23187-stimulated human mast cells (HMC-1). In addition, we showed that anti-inflammatory mechanism of AR is through suppression of nuclear factor-${\kappa}B$ activation $I{\kappa}B-{\alpha}$degradation. These results provided new insight into the pharmacological actions of AR as a potential molecule for therapy of inflammatory allergic diseases.

The Effect of the Transcriptional Regulation of Sp1 for TGF-β1 and CTGF Expression in Scar Formation (반흔형성 과정에서 Sp1 전사인자 조절에 의한 TGF-β1 및 CTGF의 발현)

  • Park, Dong Man;Sohn, Dae Gu;Han, Ki Hwan;Lee, Sun Young;Chae, Young Mi;Chang, Young Chae;Park, Kwan Kyu
    • Archives of Plastic Surgery
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    • v.33 no.1
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    • pp.39-45
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    • 2006
  • This study is to examine the relationship between TGF-b1 expression and CTGF expression, and to evaluate the effect of Sp1 blockade on the expression of TGF-b1, CTGF and extracellular genes, clones of fibroblasts stably transfected with Sp1 decoy ODN. R-Sp1 decoy ODN was highly resistant to degradation by nucleases or serum, compared to the linear or phosphorothioated-Sp1 decoy ODN. Skin wounds were created on the back of 36 anesthetized rats. They were divided into four groups-the rats with normal skin, with wounded skin without decoy, with wounded skin injected with R-Sp1 decoy, and with wounded skin injected with mismatched R-Sp1 decoy, respectively. Skins were collected at 3rd, 5th, 7th, 14th day after wounding. Cellular RNA was extracted by RT-PCR analysis. TGF-${\beta}1$ and CTGF were deeply related with skin fibrosis during scar formation and it appeared that TGF-${\beta}1$ may cause the induction of CTGF expression. R-Sp1 decoy ODN inhibited TGF-${\beta}1$ and CTGF expression both in cultured fibroblasts and in the skin of rats. These results indicate that targeting Sp1 with R-type decoy efficiently blocks extracellular matrix gene expression, and suggest an important new therapeutic approach to control the scarring in normal wound healing and fibrotic disorders.

Induction of Apoptosis by Citri Pericarpium Methanol Extract through Reactive Oxygen Species Generation in U937 Human Leukemia Cells (진피 메탄올 추출물의 활성산소종 생성을 통한 인체 백혈병 세포의 apoptosis 유발)

  • Kim, Ga Hee;Lee, Moon Hee;Han, Min Ho;Park, Cheol;Hong, Su Hyun;Choi, Yung Hyun
    • Journal of Life Science
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    • v.23 no.8
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    • pp.1057-1063
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    • 2013
  • Citri Pericarpium is one of the most commonly used traditional herbal medicines in Korea, China, and Japan. Its extracts have many properties including the treatment of indigestion and inflammatory respiratory syndromes such as bronchitis and asthma. However, the underlying molecular mechanisms of anti-cancer activity and molecular targets are not fully understood. In this work, we investigated the anti-proliferative activity of Citri Pericapium (EMCP) methanol extract on reactive oxygen species (ROS) production and the association of these effects with apoptotic cell death using U937 human leukemia cells in vitro. EMCP treatment decreased cell proliferation in a dose-dependent manner following an increase of the sub-G1 phase, the down-regulation of Bax proteins, the activation of caspases, the degradation of poly (ADP-ribose) polymerase proteins (PARP), and the induction of ROS generation. However, the quenching of ROS generation by N-acetyl-L-cysteine administration, a scavenger of ROS, reversed the EMCP-induced apoptosis effects. In addition, heme oxygenase-1 expression also recovered by inhibiting the nuclear translocation of phosphorylated NF-E2-related factor 2. Taken together, our data indicate that ROS are involved as key mediators in the early molecular events in the EMCP-induced apoptotic pathway.

A Study on the Classifying Quality Standard by Comparison with Physicochemical Characteristics of Virgin, Pure, Pomace Olive Oil (버진, 퓨어, 포마스 올리브유의 이화학적인 특성 비교를 통한 품질등급 구분에 관한 연구)

  • Cho, Eun-Ah;Lee, Young-Sang
    • The Korean Journal of Food And Nutrition
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    • v.27 no.3
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    • pp.339-347
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    • 2014
  • This study investigated the classification of olive oils that are mainly distributed in Korea via imports. The fatty acid contents, degree of color, pigments, anti-oxidants, and sterol contents are analyzed on the different types of olive oil as follows: 10 kinds of extra virgin olive oil, 5 kinds of pure olive oil, and 5 kinds of refined olive-pomace oil. As a result of fatty acid analysis, the majority of oleic acid ($C_{18:1}$) and palmitic acid ($C_{16:0}$), and minority of linoleic acid ($C_{18:2}$) and stearic acid ($C_{18:0}$) were detected without any significant differences between the grades of olive oils. The UV spectrum is related to the ${\Delta}K$, and it is a part of the analysis factor for the purity and degree of degradation of the oil. Extra virgin olive oil had ${\Delta}K$ of almost 0, pure olive oil had 0.07~0.12, and refined olive-pomace oil had 0.1~0.13. These differed from extra virgin oil, and the pure or pomace oil ${\Delta}K$ had a confirmed distinct difference. The color degrees of chlorophyll with a low $L^*$ value and $(-)a^*$ (green) and carotenoid with $(+)b^*$ (yellow) were confirmed to have correlation between extra virgin and other olive oils. To compare chlorophyll and carotenoid as natural pigment in olive oils, 417 nm and the ratio of the absorbance at 480 nm (417/480) was calculated at 1.62 of extra virgin, 1.85 of pure olive oil, and 3.32 of refined olive-pomace oil. Therefore, it will be possible to distinguish when the extra virgin or pure olive oil are mixed with olive-pomace oil. The total amount of tocopherol, an anti-oxidant, were 19.06 in extra virgin, 10.91 in pure olive oil, and 27.88 in refined olive-pomace oil. The high content of tocopherol in pomace oil caused recovery of solvent extraction from olive pulp. Thus, extra virgin oil and pure olive oil were distinguished by olive-pomace oil. Polyphenol compounds in extra virgin olive oil measured high only in ferulic acid with 0.543 mg/kg, caffeic acid with 0.393 mg/kg, and other vanillic acid, vanillin, and p-coumaric acid had similar amount of 0.3 mg/kg. All grade of olive oils had the highest ${\beta}$-sitosterol content. Af (Authenticity factor) value were estimated with campesterol and stigmasterol content ratio (%). Af value was 19.2 in extra virgin olive oil, 17.1 in pure olive oil, 16.9 in refined olive-pomace oil, which were distinctive from sunflower oil with 3.7, corn oil with 2.4, and soybean oil with 2.0. It can provide important indicator of olive oil adulteration with other cheap vegetable oils. The results of this study can be used as a database for the classification of olive oil grade and distinguishing between the different types of oils.

Studies of Inhibitory Mechanism on Melanogenesis by Partially Purified Asiasari radix in α-MSH Stimulated B16F10 Melanoma Cells (세신추출물이 α-MSH 자극에 의한 B16F10 세포의 멜라닌생성에 미치는 영향)

  • Jang, Ji-Yeon;Kim, Ha-Neui;Kim, Yu-Ri;Kim, Byung-Woo;Choi, Yung-Hyun;Choi, Byung-Tae
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1617-1624
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    • 2010
  • Recently, it has been found that Asiasari radix showed a hypopigmenting effect on melanogenesis through activation of mitogen-activated protein kinase (MEK)/extracellular signal-activated kinase (ERK) in B16F10 melanoma cells. However, the hypopigmenting effect of A. radix on the $\alpha$-melanocyte stimulating hormone ($\alpha$-MSH)-stimulated melanogenesis has remained unknown. The purpose of this study was to investigate the inhibitory mechanism of the partially purified A. radix (PPAR)-induced hypopigmentating effects on $\alpha$-MSH-stimulated melanogenesis in B16F10 mouse melanoma cells. PPAR strongly inhibited tyrosinase activity and leads to decreased melanin synthesis in $\alpha$-MSH-stimulated B16F10 melanoma cells. PPAR also decreased the $\alpha$-MSH-induced over-expression of the melanogenic enzymes, tyrosinase, tyrosinase-related protein (TRP)-1, dopachrome tautomerase (Dct) and microphthalmia-associated transcription factor (MITF). We further showed that PPAR inhibits $\alpha$-MSH-induced melanogenesis via phosphorylation of MEK/ERK and PI3K/Akt, and that their activation was blocked by MEK inhibitors, PD98059 and PI3K inhibitors, LY294002 in $\alpha$-MSH-stimulated B16F10 melanoma cells. These results suggest that PPAR inhibits $\alpha$-MSH-induced melanogenesis by activation of MEK/ERK and PI3K/Akt through MITF degradation, which may lead to down-regulation of tyrosinase.

Flexural Behaviors of GFRP Rebars Reinforced Concrete Beam under Accelerated Aging Environments (GFRP Rebar 보강 콘크리트 보의 급속노화환경에서의 휨 거동에 관한 연구)

  • Park, Yeon-Ho;Choi, Yeol
    • Journal of the Korea Concrete Institute
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    • v.25 no.2
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    • pp.137-144
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    • 2013
  • The use of fiber-reinforced polymer (FRP) reinforcing bars in concrete structures has been increased as an alternative of steel reinforcement which has shown greater vulnerability to corrosion problem. However, the long-term performance of concrete members with FRP reinforcement is still questioned in comparison to the used of steel reinforcement. This study presents the results of an experimental study on the long-term behaviors of GFRP (glass fiber reinforced polymer) bar reinforced concrete beams after exposed to accelerated aging in an environmental chamber with temperature of $46^{\circ}C$ ($115^{\circ}F$) and 80% of relative humidity up to 300 days. The objectives of this research was to compare strength degradation and change of ductility between GFRP reinforced concrete beams and steel reinforcement beams after accelerated aging. Two types (wrapped and sand-coated surface) of GFRP bars and steel were reinforced. in concrete beams. Test results show that the failure modes of GFRP bar reinforced concrete beams are very similar with traditional RC beams, and the change of load-carrying capacity of steel reinforcing concrete beam is greater than that of GFRP bar reinforcing concrete beam under the accelerated aging. Test result also shows that the use of GFRP reinforcing in concrete could be introduced more brittle failure than that of steel reinforcing for practical application. The deformability factor up to compression failures indicates no significant variation before and after exposure of accelerated aging.

Samsoeum inhibits systemic anaphylaxis and release of histamine, cytokine in vivo and in vitro

  • Kim, Su-Jin;Kim, Na-Hyung;Moon, Phil-Dong;Myung, Noh-Yil;Kim, Min-Chol;Lee, Ki-Taek;Jo, Hyung-Mook;Kim, Na-Hyun;Rim, Hong-Kun;Seo, Min-Jun;Kim, Jin-Man;Lee, Seung-Eun;An, Nyeon-Hyung;Lee, Kang-Min;Lee, Si-Hyung;Park, Yun-Jum;Jeong, Hyun-Ja;Um, Jae-Young;Kim, Hyung-Min;Hong, Seung-Heon
    • Advances in Traditional Medicine
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    • v.9 no.2
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    • pp.115-127
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    • 2009
  • Samsoeum (SSE) is used in traditional oriental medicine for various medicinal purposes. However, the exact mechanism that accounts for the anti-allergy and anti-inflammatory effects of the SSE is still not fully understood. The aim of the present study is to elucidate whether and how SSE modulates the allergic reactions in vivo, and inflammatory reaction in vitro. In this study, we showed that SSE significantly decreased compound 48/80-induced systemic anaphylaxis, ear-swelling response, histamine release from preparation of rat peritoneal mast cells and anti-dinitropheny IgE-induced passive cutaneous reaction. Also, SSE inhibited the expression of inflammatory cytokine and cyclooxygenase-2 in PMA plus A23187-stimulated human mast cells (HMC-1). In addition, we showed that anti-inflammatory mechanism of SSE is through suppression of nuclear factor-${\kappa}B$ activation and $I{\kappa}B-{\alpha}$ phosphorylation/degradation in HMC-1. These results provided new insight into the pharmacological actions of SSE as a potential molecule for therapy of inflammatory allergic diseases.

Expression System for Optimal Production of Xylitol Dehydrogenase (XYL2) in Saccharomyces cerevisiae (출아효모에서 xylitol dehydrogenase (XYL2)의 최적 생산을 위한 발현 시스템 구축)

  • Jung, Hoe-Myung;Kim, Yeon-Hee
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1403-1409
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    • 2017
  • In this study, the xylitol dehydrogenase (XYL2) gene was expressed in Saccharomyces cerevisiae as a host cell for ease of use in the degradation of lignocellulosic biomass (xylose). To select suitable expression systems for the S.XYL2 gene from S. cerevisiae and the P.XYL2 gene from Pichia stipitis, $pGMF{\alpha}-S.XYL2$, $pGMF{\alpha}-P.XYL2$, $pAMF{\alpha}-S.XYL2$ and $pAMF{\alpha}-P.XYL2$ plasmids with the GAL10 promoter and ADH1 promoter, respectively, were constructed. The mating factor ${\alpha}$ ($MF{\alpha}$) signal sequence was also connected to each promoter to allow secretion. Each plasmid was transformed into S. cerevisiae $SEY2102{\Delta}trp1$ strain and the xylitol dehydrogenase activity was investigated. The GAL10 promoter proved more suitable than the ADH1 promoter for expression of the XYL2 gene, and the xylitol dehydrogenase activity from P. stipitis was twice that from S. cerevisiae. The xylitol dehydrogenase showed $NAD^+$-dependent activity and about 77% of the recombinant xylitol dehydrogenase was secreted into the periplasmic space of the $SEY2102{\Delta}trp1/pGMF{\alpha}-P.XYL2$ strain. The xylitol dehydrogenase activity was increased by up to 41% when a glucose/xylose mixture was supplied as a carbon source, rather than glucose alone. The expression system and culture conditions optimized in this study resulted in large amounts of xylitol dehydrogenase using S. cerevisiae as the host strain, indicating the potential of this expression system for use in bioethanol production and industrial applications.