• 한국식품저장유통학회지
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• 제14권6호
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• pp.655-661
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• 2007

참당귀(Angelica gigas Nakai)를 기능성 식품소재 및 의약품의 원료로 활용하기 위하여 참당귀 methanol 추출물에 대한 항산화, 항암 및 면역활성을 조사한 결과는 다음과 같다. 참당귀의 항산화력을 알아보기 위하여 0.1, 0.5 및 1 mg/mL의 농도로 당귀의 methanol 추출물을 처리한 후 수소 공여능, 환원력 및 hydroxyl radical 활성을 측정하였다. 그 활성은 모두 추출물의 농도에 의존적으로 높게 나타났으며, 특히 수소공여능은 1 mg/mL에서 대조군과 비교했을때 50% 정도의 활성을 나타내었다. 참당귀 methanol 추출물을 SW480세포에 처리한 결과 대조군에 비하여 농도 의존적으로 암세포의 성장을 억제하였으며, 암세포에 참당귀 methanol 추출물을 24시간 처리하여 hoechst 염색한 후 형광현미경으로 관찰한 결과 핵의 응축 및 DNA 분절이 관찰되었다. 또한 참당귀 methanol 추출물이 처리된 암세포에서는 대조군에 비하여 농도에 의존적으로 caspase-3의 활성이 증가하였다. 참당귀의 methanol 추출물은 생쥐의 비장으로부터 분리한 면역세포에 대하여 대조군과 비교하여 상이한 차이를 보이지 않았으며, 또한 대식세포인 RAW 264.7에서도 $NO_2$(Nitrite)를 함량이 증가되지 않았다. 따라서 이들 결과는 참당귀 methanol 추출물은 약한 항산화 활성을 가지며, 면역 활성에는 큰 관련이 없지만, SW480 암 세포에서 caspase dependent pathway에 의한 apoptosis를 유도함으로서 암세포의 사멸하는 것으로 판단되어진다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 추계학술대회
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• pp.92-92
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• 2019

Excessive or chronic inflammation contributes to the pathogenesis of many inflammatory diseases such as sepsis, rheumatoid arthritis, and ulcerative colitis. Hibiscus syriacus L. has been used as a medicinal plant in many Asian countries, even though its anti-inflammatory activity has been unclear. Therefore, we investigated the anti-inflammatory effect of anthocyanin fractions from the H. syriacus L. varieties Pulsae (PS) on the lipopolysaccharide (LPS)-induced expression of proinflammatory mediators and cytokines in RAW264.7 macrophages. PS suppressed LPS-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) secretion concomitant with downregulation of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. Furthermore, PS inhibited the production of proinflammatory cytokines such as tumor necrosis factor-alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), and IL-12 in LPS-stimulated RAW264.7 macrophages. Further study showed that PS significantly decreased LPS-induced nuclear translocation of the nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) subunits, p65 and p50. Molecular docking data showed that many anthocyanins from PS fit into the hydrophobic pocket of MD2 and bound to Toll-like receptor 4 (TLR4), indicating that PS inhibits the TLR4-MD2-mediated inflammatory signaling pathway. Especially, apigenin-7-O-glucoside most powerfully bound to MD2 and TLR4 through LYS122, LYS122, and SER127 at a distance of $2.205{\AA}$, $3.098{\AA}$, and $2.844{\AA}$ and SER441 at a distance of $2.873{\AA}$ (docking score: -8.4) through hydrogen bonding, respectively. Additionally, PS inhibited LPS-induced TLR4 dimerization/expression on the cell surface, which consequently decreased MyD88 recruitment and IRAK4 phosphorylation. PS completely blocked LPS-mediated mortality in zebrafish larvae by diminishing the recruitment of neutrophil and macrophages accompanied by low levels of proinflammatory cytokines. Taken together, our results indicate that PS attenuates LPS-mediated inflammation in both in vitro and in vivo by blocking the TLR4/MD2-MyD88/IRAK4-$NF-{\kappa}B$ axis. Therefore, PS might be used as a novel modulatory candidate for effective treatment of LPS-mediated inflammatory diseases.

• 대한한의학방제학회지
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• 제25권1호
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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.

• 분석과학
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• 제13권5호
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• pp.659-665
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• 2000

Ozone/high pH, Ozone/$H_2O_2$ 공정 및 OH라디칼 scavenging 조건인 알칼리도가 존재하는 공정에서의 오존처러 후 생성되는 과산화수소의 농도와 $UV_{254}$ 감소율의 변화를 통하여 유기물의 분해 특성을 조사하였다. 부식산의 농도, 초기 pH 및 과산화수소와 알칼리도의 농도 등을 변화시키면서 생성되는 과산화수소의 농도와 $UV_{254}$ 감소율을 측정하였다. 그 결과, 수중 부식산의 오존처리시 부식산과 오존과의 반응에 의한 과산화수소의 생성과 부식산의 분해에 따른 $UV_{254}$의 감소는 서로 다를 반응경로를 거치는 것을 확인 하였다. 생성되는 과산화수소의 농도는 용존 오존분자의 안정성이 높은 산성조건에서 높게 나타났다. 반면, $UV_{254}$의 감소율은 강알칼리성 용액이나 $H_2O_2$가 투여된 경우, 즉 ${\cdot}OH$, ${\cdot}HO_2$와 같은 라디칼이 많이 형성되는 조건에서 높았다. 이러한 결과는 부식산의 오존처리 과정에서 생성되는 과산화수소의 농도가 용존 오존분자의 직접반응에 주로 의존하는 반면, $UV_{254}$의 감소율은 오존분자의 직접반응과 더불어 라디칼과의 간접반응에 영향을 받음을 나타낸다.

• Asian-Australasian Journal of Animal Sciences
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• 제13권6호
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• pp.748-756
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• 2000

Effects of xanthine (X), xanthine oxidase (XO), and catalase (C), $H_2O_2$, and carbohydrates on sperm capacitation, acrosome reaction, and fertilizing ability in vitro were examined. Glucose alone, but not fructose, supported the maximum rate of sperm capacitation and acrosome reaction. However, in the combination of X, XO, and C (XXOC) or $H_2O_2$, fructose alone also supported maximum capacitation, acrosome reaction, and fertilization. Either insufficient or excessive amounts of $H_2O_2$ decreased sperm capacitation and the acrosome reaction. In order to understand how glucose generates $H_2O_2$ or other reactive oxygen species in sperm cells, 6-aminonicotinamide, an inhibitor of the pentose-phosphate pathway (PPP), and apocynin, an inhibitor of NADPH oxidase, were added to sperm suspensions in glucose-containing medium. Results appeared that sperm capacitation, acrosome reaction, and fertilization were consequently inhibited by either one of these compounds. These inhibitory effects were nullified by addition of XXOC. These results support the hypothesis that glucose, in addition to being a substrate for glycolysis, facilitates sperm capacitation and the acrosome reaction by generating reactive oxygen species through G-6-P dehydrogenase and NADPH oxidase.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.78.2-79
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• 2003

Pepper defensin ( CADEFl) clone was isolated from cDNA library constructed from pepper leaves infected with avirulent strain Bv5-4a of Xanthomonu campestris pv. vesicatoria. The deduced amino acid sequence of CADEFl is 82-64％ identical to that of other plant defensins. Putative protein encoded by CADEFl gene consists of 78 amino acids and 8 conserved cysteine residues to form four structure-stabilizing disulfide bridges. Transcription of the CADEF1 gene was earlier and stronger induced by X campestris pv. vesicatoria infection in the incompatible than in the compatible interaction. CADEF1 mRNA was constitutively expressed in stem, root and green fruit of pepper. Transcripts of CADEFl gene drastically accumulated in pepper leaf tissues treated With Salicylic acid (SA), methyl jasmonate (MeJA), abscisic acid (ABA), hydrogen Peroxide (H$_2$O$_2$), benzothiadiazole (BTH) and DL-${\beta}$-amino-n-butyric acid (BABA). In situ hybridization results revealed that CADEF1 mRNA was localized in the phloem areas of vascular bundles in leaf tissues treated with exogenous SA, MeJA and ABA. Strong accumulation of CADEF1 mRNA occurred in pepper leaves in response to wounding, high salinity and drought stress. These results suggest that bacterial pathogen infection, abiotic elicitors and some environmental stresses may play a significant role in signal transduction pathway for CADEF1 gene expression.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2005년도 춘계학술대회
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• pp.53-60
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• 2005

Parkinson's disease (PD) is a progressive neurodegenerative disorder affecting $1\%$ of the population above the age of 65 and is characterized by a selective loss of dopaminergic neurons in the substantia nigra pars compacta. Although the underlying cause of dopaminergic cell death or the mechanism by which these cells degenerate is still not clearly understood, oxidative stress, mitochondrial dysfunction, and protein misfolding are thought to play important roles in the dopaminergic degeneration in PD. Tetrahydrobiopterin (BH4) is synthesized exclusively in the monoaminergic, including dopaminergic, cells and serves as an endogenous and obligatory cofactor for syntheses of the potential oxidative stressors dopamine and nitric oxide. In addition to its contribution toward the syntheses of these two potentially toxic molecules, BH4 itself can directly generate oxidative stress. BH4 undergoes oxidation during the hydroxylation reaction as well as nonenzymatic autooxidation to produce hydrogen peroxide and superoxide radical. We have previously suggested BH4 as an endogenous molecule responsible for the dopaminergic neurodegeneration. BH4 exerts selective toxicity to dopamine-producing cells via generation of oxidative stress, mitochondrial dysfunction, and apoptosis. BH4 also induces morphological, biochemical, and behavioral characteristics associated with PD in vivo. BH4 as well as enzyme activity and gene expression of GTP cyclohydrolase I, the rate-limiting enzyme in BH4 synthesis pathway, are readily upregulated by cellular changes such as calcium influx and by various stimuli including stress situations. This points to the possibility that cellular availability of BH4 might be increased in aberrant conditions, leading to increased extracellular BH4 subsequent degeneration. The fact that BH4 is specifically and endogenously synthesized in dopaminergic cells, Is readily upregulated, and generates oxidative stress-related cell death provides physical relevance of this molecule as an attractive candidate with which to explain the mechanism of pathogenesis of PD.

• Biomolecules & Therapeutics
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• 제13권4호
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• pp.207-213
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• 2005

It has been proposed that reactive oxygen species (ROS), mainly superoxide anion ($O_2^-$) and hydrogen peroxide ($H_2O_2$), may mediate oxidative stress. Production of $H_2O_2$ during oxidative phosphorylation, inflammation, and ischemia can cause oxidative stress leading to cell death. Although glucose oxidase (GOX) in the presence of glucose continuously generates $H_2O_2$, it is not clear whether GOX produces apoptotic cell death in C6 glial cells. Thus, we investigated the mechanism by which GOX induces cell death. Cells were incubated with different concentration of GOX in the presence of glucose where cell viability, TUNEL and DNA ladder were analyzed. Results indicated that GOX exhibited cytotoxicity in a dose dependent manner by MTT assay. TUNEL positive cell and DNA laddering showed that GOX-induced cytotoxicity was due to apoptosis. Western blot analysis also showed that the cleaved caspase-3 level was detected in the GOX-treated cells at 10 mU/ml and increased dramatically at 30 mU/ml. Cleaved PARP also appeared at 10 mU/ml and lasted at 20 or 30 mU/ml of GOX. Cytochrome c level was increased by GOX dose dependently, which was contrast to Bcl-2 expression level. These results suggest that GOX induces apoptosis through caspase-3 activation, which followed by cytochrome c release from mitochondria through regulating of Bcl-2 level.

• Nutrition Research and Practice
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• 제9권2호
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• pp.123-128
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• 2015

BACKGROUND/OBJECTIVES: Natural products or active components with a protective effect against oxidative stress have attracted significant attention for prevention and treatment of degenerative disease. Oligonol is a low molecular weight polyphenol containing catechin-type monomers and oligomers derived from Litchi chinensis Sonn. We investigated the protective effect and its related mechanism of oligonol against oxidative stress. MATERIALS/METHODS: Oxidative stress in C6 glial cells was induced by hydrogen peroxide ($H_2O_2$) and the protective effects of oligonol on cell viability, nitric oxide (NO) and reactive oxygen species (ROS) synthesis, and mRNA expression related to oxidative stress were determined. RESULTS: Treatment with oligonol inhibited NO and ROS formation under cellular oxidative stress in C6 glial cells. In addition, it recovered cell viability in a dose dependent-manner. Treatment with oligonol also resulted in down-regulated mRNA expression related to oxidative stress, nuclear factor kappa-B (NF-${\kappa}B$) p65, cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS), compared with the control group treated with $H_2O_2$. In particular, expression of NF-${\kappa}B$ p65, COX-2, and iNOS was effectively reduced to the normal level by treatment with $10{\mu}g/mL$ and $25{\mu}g/mL$ of oligonol. CONCLUSIONS: These results indicate that oligonol has protective activity against oxidative stress-induced inflammation. Oligonol might be a promising agent for treatment of degenerative diseases through inhibition of ROS formation and NF-${\kappa}B$ pathway gene expression.

• 한국미생물·생명공학회지
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• 제46권2호
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• pp.145-153
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• 2018

Among the carotenoid biosynthesis genes, crtI gene encodes the phytoene desaturase (CrtI) enzyme, and phytoene desaturase convert phytoene to lycopene. Phytoene desaturase is involved in the dehydrogenation reaction, in which four single bonds in the phytoene are introduced into a double bond, eliminating eight hydrogen atoms in the process. Phytoene desaturase is one of the key regulating enzyme in carotenoid biosynthetic pathway of various carotenoid biosynthetic organisms. The crtI gene in genomic DNA of Paracoccus haeundaensis was amplified and cloned into a T-vector to analyze the nucleotide sequence. As a result, the crtI gene coding for phytoene desaturase from P. haeundaensis consists of 1,503 base pairs encoding 501 amino acids residues. An expression plasmid containing the crtI gene was constructed, and Escherichia coli cells containing this plasmid produced the recombinant protein of approximately 55 kDa, equivalent to the molecular weight of phytoene desaturase. The expressed protein in cell lysate showed enzymatic activity similar to phytoene desaturase. Phytoene and lycopene were analyzed by HPLC and measured at wavelength of 280 nm and 470 nm, respectively. The $K_m$ values for phytoene and NADPH were $11.1{\mu}M$ and $129.3{\mu}M$, respectively.