• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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• pp.176-176
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• 1998

An enhanced formation of nitric oxide (NO) is an important mediator of hypotention, peripheral vasodilation and vascular hyporeactivity to vasoconstrictor agents in endotoxaemia. And tumor necrosis factor (TNF${\alpha}$), as a primary mediator of circulatory shock has been known to induce inducible nitric oxide synthase (i-NOS), leading to excessive production of NO. We isolated two sesquiterpene lactone compounds from Saussurea lappa and their structures were elucidated as dehydrocostus lactone and costunolide. These compounds inhibit the production of both NO and TNF${\alpha}$ by LPS (1 $\mu\textrm{g}$/$m\ell$)-activated Raw 264.7 cells. NO was measured spectropho-tometrically as nitrite by the Griess reagent and TNF${\alpha}$ by ELISA. Dehydrocostus lactone (IC$\sub$50/ : 3.0 ${\mu}$M) and costunolide (IC$\sub$50/ : 4.5 ${\mu}$M) inhibited the production of NO in LPS-activated Raw 264.7 cells by suppressing inducible nitric oxide synthase enzyme expression. These compounds also decreased the TNF${\alpha}$ levels in LPS-activated system in vitro and in vivo.

• 대한본초학회지
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• 제36권6호
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• pp.17-25
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• 2021

Objectives : Baicalein (3,3', 4', 5, 6-pentahydroxyflavone), a type of flavonoid, is a well-known antioxidant and anti-inflammatory ingredient found in Scutellaria baicalensis root. The aim of this study is to investigate the effect of baicalein on nitric oxide (NO) production in TM3 mouse Leydig cells stimulated by indomethacin (IN). Methods : TM3 cells were treated with IN (0.5 μM) and baicalein at concentrations of 12.5, 25, 50, and 100 μM for 24 hr, 40 hr, 42 hr, 44 hr, and 64 hr. After treatments, cell viabilities were measured with the modified MTT assay. The production of nitric oxide in cells was measured by Griess reagent assay. Results : Baicalein showed no cytotoxicity on IN-stimulated TM3. NO production in IN-stimulated TM3 treated for 24 hr with baicalein at concentrations of 12.5, 25, 50, and 100 μM was 95.8%, 94.86%, 89.97%, and 81.52% of the control group treated with IN only, respectively; NO production for 40 hr was 97.34%, 97.34%, 95.15%, and 87.42%, respectively; NO production for 42 hr was 89.12%, 90.14%, 89.74%, and 90.26%, respectively; NO production for 44 hr was 83.83%, 84.94%, 85.65%, and 86.85%, respectively; NO production for 64 hr was 94.12%, 95.38%, 94.21%, and 94.12%, respectively. Specifically, baicalein at concentrations of 12.5, 25, and 50 have been shown to most efficiently inhibit NO productions in 48 hr of treatment. Conclusions : Baicalein might have anti-toxicant effect on Leydig cells related with its inhibition of NO production in Leydig cells stimulated with IN.

• The Korean Journal of Physiology and Pharmacology
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• 제10권3호
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• pp.155-159
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• 2006

Among the Shc proteins, p66shc is known to be related to oxidative stress responses and regulation of the production of reactive oxygen species (ROS). The present study was undertaken to investigate the role of p66shc on endothelial nitric oxide synthase (eNOS) activity in the mouse embryonic fibroblasts (MEFs). When wild type (WT) or p66shc (-/-) MEFs were transfected with full length of eNOS cDNA, the expression and activity of eNOS protein were higher in the p66shc (-/-) MEFs. These phenomena were reversed by reconstitution of p66shc cDNA transfection in the p66shc (-/-) MEFs. The basal superoxide production in the p66shc (-/-) MEFs was not significantly different from that of WT of MEFs. However, superoxide production induced by NADPH in the p66shc (-/-) MEF was lesser than that in WT MEFs. When compared with WT MEFs, cell lysate of p66shc (-/-) MEFs showed significantly increased H-ras activity without change of endogenous H-ras expression. Our findings suggest the pivotal role of p66shc adaptor protein played in inhibition of endothelial nitric oxide production via modulation of the expression and/or activity of eNOS protein.

• 동의생리병리학회지
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• 제17권3호
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• pp.771-776
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• 2003

The present study was conducted to evaluate the regulation mechanism of nitric oxide(NO) by water-extracted Cyperus rotundus (WCR) in RAW 264.7 macrophages. We investigated the effects of cell proliferation in mouse spleen cell and RAW 264.7 macrophages cells. WCR enhanced mitogenic activity in the dose-response manner in mouse spleen cells and RAW 264.7 macrophages cells. In nitric oxide (NO) synthesis by WCR, WCR alone had an effect on NO synthesis. It was found that the production of NO of RAW 264.7 cells stimulated with lipopolysaccharide (LPS) could be markedly inhibited by WCR. Inhibition of NO production was achieved by reducing inducible nitric oxide syntheses(iNOS) mRNA expression. The expression of IL-I gene by WCR was investigated using reverse transcription polymerase chain reaction (RT-PCR). In RT-PCR, IL-1 family(IL-1 α, IL-1β) expressions were induced by WCR. These finding suggested that regulation of NO production by WCR may be, at least in part, associated with the regulation of iNOS mRNA expression and IL-1 family gene expression.

• 한국식품영양과학회지
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• 제42권9호
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• pp.1513-1517
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• 2013

본 연구는 사과씨로부터 추출한 사과씨 에탄올 추출물이 1차 면역세포인 대식세포의 면역기능에 관하여 면역기능을 증가시켜 줄 수 있는 지에 관한 여부를 알아보기 위하여 수행되었다. 사과씨 추출물을 마우스 유래의 대식세포인 RAW 264.7 세포에 처리하였을 때, 대식세포의 활성화 관련 지표인 nitric oxide와 cytokine(IL-6, TNF-${\alpha}$)의 생성이 증가되었다. 이러한 결과는 사과씨 추출물이 대식세포의 활성에 크게 영향을 줄 수 있다는 가능성을 제시하고, 이러한 사과씨 추출물이 대식세포의 면역 활성을 유도하는 신호전달 과정에 관하여 연구해 본 결과, 사과씨 추출물의 처리는 대식세포 내 MAPKs(ERK, p38) 및 $I{\kappa}B-{\alpha}$의 인산화를 증가시키는 신호전달 과정을 경유하는 것으로 관찰되었다.

• 생약학회지
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• 제36권3호통권142호
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• pp.245-251
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• 2005

Phytochemical examination of the barks of Actinidia arguta led to the isolation of five flavonoids. Structures of compounds were elucidated as catechin (1), (-)-epicatechin (2), quercetin (3), $quercetin-3-O-{\beta}-D-glucopyranoside$ (4), $quercetin-3-O-{\beta}-D-galactopyranoside$ (5) by comparison with previously reported spectral evidences. To investigate the anti-oxidative effect and nitric oxide (NO) production inhibitory activity of these compounds, DPPH radical scavenging activity and nitric oxide production inhibitory activity in $IFN-{\gamma}$, LPS stimulated RAW 264.7 cell were examined. The $IC_{50}s$ were determinied as follows : $1\;$IC_{50}=26.61\;{\mu}g/ml$, $2\;IC_{50}=25.30\;{\mu}g/ml$, $3\;IC_{50}=20.41\;{\mu}g/ml$, $4\;IC_{50}=18.23\;{\mu}g/ml$ , $5\;IC_{50}=30.46\;{\mu}g/ml$, $6\;IC_{50}=28.0;{\mu}g/ml$, $7\;IC_{50}=27.24\;{\mu}/ml$. These NO production inhibitory effects were significantly different compared with the positive control, L-NMMA $(IC_{50}=20.77\;{\mu}g/ml)$, respectively. Compound $1\;(IC_{50}=6.19\;{\mu}g/ml)$, $2\;(IC_{50}=8.98\;{\mu}g/ml)$, $3\;(IC_{50}=7.30\;{\mu}g/ml)$ and $4\;(IC_{50}=7.64\;{\mu}g/ml)$ also showed potent antioxidative activities similar level to ascorbic acid $(IC_{50}=9.22\;{\mu}g/ml)$. These results suggest that barks of A. arguta have a potent anti-oxidative and anti-inflammatory activity.

• Biomolecules & Therapeutics
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• 제8권2호
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• pp.140-146
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• 2000

Recently, we reported that immunostimulation of primary rat cortical astrocyte caused stimulation of glucose deprivation induced apoptotic cell death. To enhance the understanding of the mechanism of the potentiated cell death of clucose-deprived astrocyte by immunostimulation, we investigated the effect of immunostimulation on the glucose deprivation induced cell death of rat C6 glioma cells. Co-treatment of C6 glioma cells with lipopolysaccharide (LPS, $1\;{\mu}\textrm{g}/ml$) and interferon ${\gamma}(IFN{\gamma},\;100U/ml)$ is serum free condition caused marked elevationo f nitric oxide production ($>50\;{\mu}M$). In this condition, glucose deprivation caused significant release of lactate dehdrogenase (LDH) from C6 glioma cells while control cells did not show LDH release. To investigate whether elevated level of nitric oxide is responsible for the enhanced LDH release in glucose-deprived condition, C6 glioma cells were treated with 3-morphorinosydnonimine (SIN-1) and it was observed that SIN-1 caused increase in LDH release from glucose-deprived C6 glioma cells. Treatment of C6 glioma cells with $25\;{\mu}M$ of pyrrolidinedithiocarbamate (PDTC) which inhibit Nuclear factor kB (NF-kB) activation, caused complete inhibition of nitric oxide production. Treatment of C6 glioma cells with NO synthase inhibitors, $N^{G}$-nitro-L-arginine (NNA) or L-$N{\omega}$-nitro-L-arginine methyl ester (L-NAME), caused inhibition of nitric oxide production and also glucose deprivation induced cell death of cytokine-stimulated C6 glioma cells. In addition, diaminohydroxypyrimidine (DAHP, 5 mM) which inhibits the synthesis of tetrahydrobiopterine (BH4), one of essential cofactors for iNOS activity, caused complete inhibition of NO production from immunostimulated C6 glioma cells. The results from the present study suggest that immunostimulation causes potentiation of glucose deprivation induced death of C6 glioma cells which is mediated at least in part by the increased production of nitric oxide. The vulnerability of immunostimulated C6 glioma cells to hypoglycemic insults may implicate that the elevated level of cytokines in various ischemic and neurodegenerative diseases may play a role in their pathogenesis.

• Archives of Pharmacal Research
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• 제23권1호
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• pp.54-58
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• 2000

To elucidate the molecular mechanisms for the suppression of LPS-induced nitric oxide (NO) production by a dehydrocostus lactone (DL) from Saussurea lappa, we examined the preventive effect of this compound on $NF-{\kappa}B$ activation in LPS-treated RAW 264.7 macrophages and U937 human monocytic cells. The results suggest that the suppression of NO production is mediated by the inhibitory action on the i-NOS gene expression through the inactivation of $NF-{\kappa}B$ and this sesquiterpene lactone can act as a pharmacological inhibitor of the $NF-{\kappa}B$ activation.

• 셀메드
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• 제8권2호
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• pp.8.1-8.5
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• 2018

Nitric oxide (NO) is a small diffusible molecule which plays an important role in various physiological activities. NO is a notable molecule, functioning as a cytotoxic agent and cellular messenger. There has been considerable interest in NO production by activated macrophages because this gaseous metabolite plays a fundamental role in the cytotoxic and cytostatic effects of macrophages towards invasive micro-organisms and tumour cells. No is a bioactive free radical that has been implicated in many physiological functions, plays a critical role during inflammation and therefore constitutes a potential target for developing therapeutics for inflammatory diseases. The use of medicinal plants by the population has been an important alternative the resource in the treatment of various diseases. Its growing acceptance in the medical community has been due to the fact that several plants with biological activities have been scientifically investigated and their efficacy and safety have been proven. In this review, discussed suppressive effects of No production by traditional medicines in RAW 264.7 and THP-1 macrophages.

• 대한한의학방제학회지
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• 제6권1호
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• pp.175-186
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• 1998

The purpose of this research was to investigate effects of Astragali Radix(AR) and Armeniacae Semen(AS) on T-lymphocytes and peritoneal macrophages in mice. The proliferation of thymocytes and splenocytes were teated using macroplate-reader. The apoptosis and sub-population of T-lymphocytes were tested using a flow cytometer. Nitric oxide production was tested using a Griess reagents. The result were obtained as follow; 1. AR increased the proliferation of thymocytes and splenocytes. 2. AS decreased the proliferation of thymocytes and splenocytes. 3. AR and AS decreased No production fron peritoneal macrophages 4. AR and AS were accelerate T-lymphocytes apoptosis. 5. AR and AS increased $T_C$ cells population, but decreased $T_H$ cells population of T-lymphocyte.