• Biomolecules & Therapeutics
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• 제21권1호
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• pp.42-48
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• 2013

Nystatin, a polyene antifungal antibiotic, is a cholesterol sequestering agent. The antifungal agent alters composition of the plasma membrane of eukaryotic cells, whereas its effects on cells are poorly investigated. In the current study, we investigated the question of whether nystatin was able to induce expression of macrophage inflammatory protein-1 (MIP-1). THP-1 cells rarely express MIP-$1{\alpha}$ and MIP-$1{\beta}$, however, upon exposure to nystatin, significantly elevated expression of MIP-$1{\alpha}$ and MIP-$1{\beta}$ was observed in a dose-dependent fashion at the messenger and protein levels. Cellular factors activated by nystatin as well as involved in nystatin-induced expression of MIP-1 proteins were identified in order to understand the molecular mechanisms of action of the anti-fungal agent. Treatment with nystatin resulted in enhanced phosphorylation of Akt, ERK, p38 MAPK, and JNK. Abrogation or significant attenuation of nystatin-induced expression of MIP-$1{\alpha}$ and MIP-$1{\beta}$ was observed by treatment with Akt inhibitor IV, LY294002, and SP6001250. Inhibition of ERK or p38MAPK using U0126 and SB202190 did not lead to attenuation of MIP-1 expression. In addition, inhibitors of protein kinase C, such as GF109203X and Ro-318220, also attenuated expression of MIP-1. These results indicate that nystatin is able to activate multiple cellular kinases and, among them, Akt and JNK play primary roles in nystatin-induced expression of MIP-1 proteins.

• Natural Product Sciences
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• 제23권3호
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• pp.162-168
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• 2017

Cordyceps bassiana has long been used as an oriental medicine and reported to possess diverse biological activities. The fruiting bodies of Cordyceps bassiana was extracted with ethanol and then further fractionated with n-hexane, ethyl acetate, n-butanol and water. The butanol fraction from Cordyceps bassiana (CBBF) exhibited the most effective in anti-inflammatory activity in RAW 264.7 macrophages and the roles of CBBF on the anti-inflammation cascade in LPS-stimulated RAW 264.7 cells were studied. To investigate the mechanism by which CBBF inhibits NO, iNOS and COX-2, the activation of $I{\kappa}B$ and MAPKs in LPS-activated macrophage were examined. Our present results demonstrated that CBBF inhibits NO production and iNOS expression in LPS-stimulated RAW 264.7 macrophage cells, and these effects were mediated through the inhibition of $I{\kappa}B-{\alpha}$, JNK and p38 phosphorylation. Also, CBBF suppressed activation of MAPKs including p38 and SAPK/JNK. Furthermore, CBBF significantly suppressed LPS-induced intracellular ROS generation. Its inhibition on iNOS expression, together with its antioxidant activity, may support its anti-inflammatory activity. Thus Cordyceps bassiana can be used as a useful medicinal food or drug for further studies.

• 한국식품과학회지
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• 제36권6호
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• pp.986-990
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• 2004

대식세포는 IL-1, IL-6, $TNF-{\alpha},\;IFN-{\gamma}$의 중요한 근원이고, 전염성의 병원체와 종양 세포에 대해 저항하는 주요한 effector 세포이다. 이것은 cytokine signal에 의해 세포파괴가 되도록 활성화 된다. 이번 연구에서 우리는 산더덕의 물 추출물이 cytokine, nitric oxide와 같은 effector 분자를 유도하기 위해 대식세포를 활성화하는지를 알아보기 위하여 실험하였다. 산더덕 추출물에 의한 대식세포의 NO 생성은 농도 의존적이고, 시간 의존적이었다. iNOS는 시간이 지날수록 발현이 유도되었다. 산더덕 추출물에 의한 IL-1과 IL-6 mRNA 유도는 시간 의존적으로 증가 되었고, 처리 후 24시간에 최고점에 도달하였다. 이것은 활성화된 대식세포가 종양세포를 죽일 수 있음을 말한다. $TNF-{\alpha}\;mRNA$의 양은 시간이 지나도 일정하였고, $IFN-{\gamma}\;mRNA$의 양은 산더덕 추출물의 처리 1시간 후에 빠르게 강화되었다. 이러한 결과로부터 산더덕은 효과적인 면역조절자이고 대식세포의 항종양활성을 강화함을 알 수 있다.

• 동의생리병리학회지
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• 제20권3호
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• pp.603-608
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• 2006

In activated macrophage, large amounts of nitric oxide (NO) are generated by inducible nitric oxide synthase (iNOS), resulting in acute or chronic inflammatory disorders. In Raw 264.7 cells stimulated with lipopolysaccharide (LPS) to mimic inflammation, water extracts from the fruit of Chaenomeles sinensis, the root of Polygonum cuspidatum and Boswellia carterii inhibited the LPS-induced NO production in a parallel dose-dependent manner. To investigate the mechanism by which those extracts inhibits NO production, we examined the expression of iNOS and COX-2 in both mRNA and protein levels. We observed a significant change in the iNOS expression between LPS alone and LPS plus those extracts-treated cells. However, water extracts from Chaenomeles sinensis, Polygonum cuspidatum and Boswellia carterii did not inhibit COX-2 expression which was induced by LPS treatment in Raw 264.7 cells. These data suggest that water extracts from Chaenomeles sinensis, Polygonum cuspidatum and Boswellia carterii can modulate anti-inflammatory immune response, which may be in part associated with the regulation of NO synthesis through the regulation of iNOS expression in mouse macrophage cells.

• Bulletin of the Korean Chemical Society
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• 제35권10호
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• pp.2945-2949
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• 2014

As a part of ongoing phytochemical research on Euonymus alatus leaves and twigs, we have isolated one new reissantane-type triterpene (1), together with eight known triterpenes, isoarborinol (2), friedelin (3), abruslactone A (4), $3{\beta},22{\alpha}$-dihydroxyolean-12-en-29-oic acid (5), $3{\alpha},22{\alpha}$-dihydroxyolean-12-en-29-oic acid (6), $3{\alpha},22{\beta}$-dihydroxyolean-12-en-29-oic acid (7), $22{\alpha}$-hydroxy-3-oxoolean-12-en-29-oic acid (8), demethylregelin (9). The structure of 1 has been elucidated as 24,24-dimethyl-reissant-7(8),25-dien-$3{\alpha}$-ol, by extensive 1D and 2D spectroscopic methods including $^1H$-NMR, $^{13}C$-NMR, $^1H-^1H$ COSY, HSQC, HMBC and NOESY. Anti-inflammatory activities of the isolated compounds were determined as potential regulating excessive inflammatory responses in RAW264.7 macrophage cells. Particularly, abruslactone A (4) and demethylregelin (9) showed the most potent activity, which effectively reduced the expression of iNOS protein and subsequent nitric oxide production induced by lipopolysaccharide in RAW264.7 cells.

• Journal of Ginseng Research
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• 제38권3호
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• pp.208-214
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• 2014

Background: In previous work, we reported that Korean Red Ginseng saponin fraction (RGSF) showed anti-inflammatory activities in vitro and in vivo. Methods: The present study investigated the radioprotective properties of RGSF by examining its effects on ionizing radiation (IR)-enhanced and lipopolysaccharide (LPS)-mediated inflammatory responses in murine macrophage cells. Results: RGSF induced strong downregulation of IR-enhanced and LPS-induced proinflammatory responses such as nitric oxide (NO) production (Inhibitory Concentration $50(IC_{50})=5.1{\pm}0.8{\mu}M$) and interleukin-$1{\beta}$ levels. RGSF was found to exert its radioprotective effects by inhibition of a signaling cascade that activated checkpoint kinase 2enuclear factor-${\kappa}B$. In addition, RGSF strongly inhibited IR-enhanced LPS-induced expression of hemoxyganase-1, implying that the latter may be a potential target of RGSF. Conclusion: Taken together, our data suggest that RGSF can be considered and developed for use as an effective radioprotective agent with minimal adverse effects.

• 약학회지
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• 제47권4호
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• pp.234-238
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• 2003

Linarin is a main compound from Chrysanthemum zawadskii var, latilobum. However, the biological mechanisms of these activities are unclear. Because of this wide diversity of effects, it is believed that they may be exerted through pluripotent effectors of linarin. In our previous screening study, the effects of linarin on the mouse macrophages cell line, RAW 264.7 cells, were investigated. It was found that linarin could stimulate macrophages activation by the production of tumor necrosis factor (TNF). The linarin (6.25∼12.5 $\mu\textrm{g}$/mι) inhibited the production of NO in LPS-activated RAW 264.7 cells and linarin became an useful candidates for the development of new drug to treat endotoxemia and inflammation accompanied by the overproduction of NO. However, linarin-treated total lymphocyte showed cytotoxicity in a dose dependent manner between 20 $\mu\textrm{g}$/mι and 40 $\mu\textrm{g}$/mι. In this study, linarin derivative (acetylated linarin) was synthesized in order to obtain less-cytotoxicity of linarin and evaluated for their in vitro cytotoxic activity aganist mouse total lymphocyte. There was no cytotoxic activity in a dose dependent manner (20∼40 $\mu\textrm{g}$/mι) of acetylated linarin whereas linarin showed. The production of NO, however, was not the case by this modified linarin. The cell morphological change was not significantly changed in response to acetylated linarin alone and these effects were potentiated by the addition of LPS. These results suggest that acetylated linarin may be developed to be a promising new drug candidate without cytotoxicity on the basis of its activity of macrophage activation.

• Journal of Ginseng Research
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• 제30권4호
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• pp.181-187
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• 2006

Protopanaxadiol (PPD) is a mixture of protopanaxadiol type saponins with a dammarane skeleton, from Korean red ginseng (Panax ginseng C.A. Meyer; Araliaceae). Korean ginseng is well-known herb to treat almost all kinds of diseases in Oriental medicine. This herb was particularly prescribed for treatment various inflammatory diseases, including rheumatoid arthritis, atherosclerosis, and diabetes mellitus, for centuries. To understand the efficacy of ginseng against inflammatory diseases, we aimed to show anti-inflammatory activities of the PPD in murine macrophage cell line, RAW264.7 cells using nitric oxide (NO) production assay and the expressions of pro-inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), and IL-6, and monocyte chemotactic protein-1 (MCP-1). We found that PPD saponin significantly blocked LPS ($1{\mu}g/ml$)-induced NO production in a dose-dependent manner. In addition, PPD abrogated the expressions of LPS-induced pro-inflammatory cytokines, such as IL-$1{\beta}$ and MCP-1. Moreover, cyclooxygenase (COX)-2, a critical enzyme to produce prostaglandin E2 (PGE2), was significantly inhibited by PPD in LPS-activated RAW264.7 cells. Taken together, these results suggested that anti-inflammatory efficacy of Korean red ginseng on inflammatory diseases is, at least, due to the NO inhibitory activity and the inhibition of the expressional level of inflammatory cytokines and/or mediators.

• 한국식품과학회지
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• 제46권4호
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• pp.511-515
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• 2014

각종 염증성 질환 및 패혈증으로 인한 치명적인 저혈압을 예방치료하는 약물 개발을 위한 기초 연구로서 유도성 NOS (inducible nitric oxide synthase, iNOS) 에 의한 NO의 과다 생성을 저해하는 성분을 천연물로부터 찾아내고자 본 연구를 수행하였다. NO 생성 저해활성의 검정은 대식세포주인 RAW 264.7 세포를 LPS로 활성화한 후, 유도되는 iNOS에 의해 생성되는 NO를 Griess 시약을 이용해 $NO_2{^-}$의 형태로 정량하였다. 또한 Western blot 실험 및 RT-PCR 실험을 시행하여 iNOS의 mRNA의 발현 및 단백 합성에 대한 영향을 조사하였다. 고량강(Alpinia officinarum Hance, Zingiberaceae)의 메탄올 추출물로부터 극성에 따른 용매 분획을 시행하여 활성성분을 분리하고 분광학적 분석법을 이용하여 분리한 단일성분이 flavonol 구조인 3,5,7-trihydroxy-2-phenylchromen-4-one (galangin, GLG)임을 확인하였다. 작용기전을 알아보기 위해, Western blot 및 RT-PCR 실험결과, 분리한 flavonol 성분(GLG)의 NO 생성저해 활성은 iNOS mRNA발현을 저해하여 iNOS 효소 단백질의 생성이 억제됨에 기인하는 것으로 확인하였다. 따라서, 고량강 추출물로부터 분리한 flavonol 화합물(GLG)이 iNOS 발현의 억제를 통해 다량의 NO 생산을 저해함으로써, 고량강(Alpinia officinarum)의 NO 과량생성과 관련된 염증성 질환에 대한 응용 가능성이 클 것으로 기대된다.

• The Korean Journal of Physiology and Pharmacology
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• 제2권3호
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• pp.377-384
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• 1998

Gold compounds depress phagocytic cell responses, including chemotaxis, and respiratory burst. However, the effects of gold compounds on the function of phagocytic cells are variable according to the preparation of medicine. In this study, effect of tetrachloroauric acid on activated neutrophil responses, including respiratory burst, lysosomal enzyme release and change of intracellular $Ca^{2+}$ level and on the synthesis of interleukin-8 and granulocyte-macrophage colony stimulating factor by macrophages was studied. This study further examines how gold compounds affect the activation processes. The respiratory burst stimulated by complement C5a, degraded IgG and PMA in neutrophils was inhibited by tetrachloroauric acid. In contrast to C5a and degraded IgG, PMA-stimulated superoxide production was weakly inhibited by tetrachloroauric acid. Staurosporine, genistein, EGTA and verapamil inhibited superoxide and $H_2O_2$ production caused by C5a and degraded IgG. PMA-stimulated superoxide production was inhibited by staurosporine but was not affected by genistein. Tetrachloroauric acid, genistein, EGTA and verapamil inhibited the release of acid phosphatase and myeloperoxidase, while the effect of staurosporine was not detected. The synthesis of interleukin-8 and granulocyte-macrophage colony stimulating factor by $interleukin-1{\beta}$ in macrophages was inhibited by tetrachloroauric acid. Preincubation with tetrachloroauric acid, genistein, EGTA and verapamil, the elevation of [$Ca^{2+}_i$] evoked by C5a was inhibited. Store-regulated $Ca^{2+}$ entry in thapsigargin-pretreated neutrophils was decreased by the addition of tetrachloroauric acid and genistein. The effect of staurosporine on intracellular $Ca^{2+}$ mobilization was not observed. In conclusion, tetrachloroauric acid may suppress neutrophil responses through its inhibitory action on elevation of intracellular $Ca^{2+}$ level and protein kinase C. It might exhibit an inhibitory effect on the action of protein tyrosine kinase. Tetrachloroauric acid depresses cytokine production by macrophages.