• 한국응용과학기술학회지
• /
• 제36권1호
• /
• pp.13-22
• /
• 2019

울금(Curcuma longa L.)의 산화억제 및 질소산화물 소거활성 등 기능성 소재로서 활용 가능성을 검토한 결과, 프로안토시아니딘(proanthocyanidin) 함량은 $69.000{\pm}2.737mg$ catechin equivalents (CE)/g dry weight으로 확인되었으며, 증류수(distilled water, DW), 70% 에탄올 및 노르말 부탄올(n-butanol)의 3가지 용매를 사용한 추출 수율은 DW (17.11%), 70% 에탄올(15.26%), 노르말 부탄올(4.12%) 순으로 관찰되었다. 총 플라보노이드(total flavonoid) 함량은 DW, 70% 에탄올 및 노르말 부탄올에서 각각 0.032, 0.512 및 2.221 mg quercetin equivalents (QE)/g의 함량으로 나타났고, 노르말 부탄올에서는 유의적인 차이를 보이며 높게 관찰되었다(p<0.05). Nitric oxide (NO) 라디칼 소거 활성은 농도 별(0.2~0.8 mg/mL) DW에서 15.64~26.20%, 70% 에탄올 10.52~20.76%, 노르말 부탄올에서 13.39~69.92%로 확인되었다. Nitrite ($NO_2$) 소거활성은 DW 및 70% 에탄올과 비교하였을 때 노르말 부탄올에서 강한 $NO_2$ 소거활성을 보였다. ${\beta}$-carotene 탈색 저해활성은 DW에서 8.81~25.93%, 70% 에탄올 1.20~20.20%, 노르말 부탄올 12.08~43.93%로 나타났다. 지질과산화 저해활성은 DW, 70% 에탄올 및 노르말 부탄올에서 각각 5.60~27.54%, 37.78~50.79% 및 41.79~46.39%로 동정되었다. 이에, 천연 항산화제 등 기능성 소재로서의 활용 가능성이 있을 것으로 사료된다.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
• /
• pp.145-145
• /
• 1998

In the present study, we have demonstrated that taurine could stimulate the production of nitric oxide and the activity of ERK2 (extracellular signal regulated protein kinase or pp42 MAP kinase). Nitric oxide(NO), the product of inducible nitric oxide synthase(iNOS), is known to be implicated in the metabolism of bone. ERK cascade plays a key role in the gene expression of iNOS in osteoblastic cell. We investigated whether taurine (l-20mM) could stimulate ERK2 activity, nitric oxide production, and inducible nitric oxide synthase in osteoblast-like UMR-106 cells. Nitric oxide was measured spectophotometrically as nitrite and the activation of ERK2 and iNOS was studied using Western 145 blot analysis. Taurine increased the production of nitric oxide in a dose-dependent manner and the effect was reached to a maximum at 10 mM. The activation of iNOS were consistent with NO levels. The tyrosine phosphorylation of ERK2 was increased by taurine in a time-dependent manner. The these result suggest that taurine might stimulate the production of nitric oxide in osteoblast-like cells by the activation of ERK2 and could regulate the metabolism of bone via nitric oxide.

• Journal of Preventive Medicine and Public Health
• /
• 제29권3호
• /
• pp.495-505
• /
• 1996

수은이 세포성 면역반응에 미치는 효과와 그 기전을 밝히기 위해 cytokine-EMT-6 cell-NO 모형에 대조군에는 수은을 첨가하지 않았고 나머지 실험군에는 $0.05-0.8{\mu}M$의 mercury chloride를 처리하였다. 그리고 배양시간별, 수은농도별로 세포생존율, 세포성 면역의 지표인 NO를 nitrite로 간접정량하고 대사과정에 필수 에너지요소인 nitrite를 측정한 결과는 아래와 같이 요약된다. 1. EMT-6세포의 세포생존율은 대조군과 36시간째 $0.8{\mu}M(89.50%)$을 제외한 모든 수은첨가군에서 모두 90%이상의 값을 나타내었으며, 배양시간 및 첨가한 수은농도별 군간 차이는 없었고 모든 수은첨가군과에서 대조군에 비해 큰 차이가 없었다. 2. 수은에 노출된 EMT-6 세포의 nitrite 생성량 및 ATP생성량은 배양시간의 경과에 따라 전반적으로 증가하였으며 시간경과에 따른 nitrite 생성량과 ATP 생성량은 첨가한 수은농도의 변화에 따라 각 군간에 현저한 차이를 보였다. 그리고 시간경과의 영향을 보정한 상태에서도 첨가한 수은농도의 변화에 따라 각 군간 차이는 모두 현저하게 나타났다. 한편, Nitrite 생성량과 ATP 생성량은 둘 다 수은첨가군 모두에서 대조군의 값에 비해 현저하게 낮았고, 첨가된 수은농도와 nitrite 생성량 및 ATP 생성량 사이에는 높은 음의 상관관계가 있어 수은농도의 증가에 따라 용량-반응의 관계(dose-dependent)로 감소하는 경향을 보였다. 이상의 결과, 배양조건에 수은의 첨가로 인하여 nitrite 생성량과 ATP 생성량이 동시에 감소하는 바, 수은에 의한 세포성 면역의 저하는 면역세포의 NO 생성량의 감소에 의한 것으로 사료되며, 이는 수은이 세포내 에너지생산에 관련된 대사과정을 억제시키므로 나타나는 결과라고 판단된다.

• The Korean Journal of Physiology and Pharmacology
• /
• 제3권4호
• /
• pp.375-382
• /
• 1999

Growing evidence indicates that enhanced generation or actions of nitric oxide (NO) are implicated in the pathogenesis of hypertension in spontaneously hypertensive rats and diabetic nephropathy in streptozotocin (STZ)-induced diabetic rats. We investigated whether inducible nitric oxide synthase (iNOS) expression in STZ-induced diabetic rats is responsible for the alterations of vascular reactivity. Diabetic state was confirmed 28 days after injection of STZ (i.p) in rats by measuring blood glucose. In order to evaluate whether short term (4 weeks) diabetic state is related with altered vascular reactivity caused by iNOS expression, isometric tension experiments were performed. In addition, plasma nitrite/nitrate (NOx) levels and expression of iNOS in the lung and aorta of control and STZ-treated rats were compared by using Griess reagent and Western analysis, respectively. Results indicated that STZ-treated rats increased the maximal contractile response of the aorta to phenylephrine (PE), and high $K^+,$ while the sensitivity remained unaltered. Endothelium-dependent relaxation, but not SNP-mediated relaxation, was reduced in STZ-treated rats. Plasma nitrite/nitrates are significantly increased in STZ-treated rats compared to controls. The malondialdehyde (MDA) contents of liver, serum, and aorta of diabetic rats were also significantly increased. Furthermore, nitrotyrosine, a specific foot print of peroxynitrite, was significantly increased in endothelial cells and smooth muscle layers in STZ-induced diabetic aorta. Taken together, the present findings indicate that enhanced release of NO by iNOS along with increased lipid peroxidation in diabetic conditions may be responsible, at least in part, for the augmented contractility, possibly through the modification of endothelial integrity or ecNOS activity of endothelium in STZ-diabetic rat aorta.

• Biomolecules & Therapeutics
• /
• 제22권5호
• /
• pp.390-399
• /
• 2014

Chalcones (1,3-diaryl-2-propen-1-ones), a flavonoid subfamily, are widely known for their anti-inflammatory properties. Propenone moiety in chalcones is known to play an important role in generating biological responses by chalcones. In the present study, we synthesized chalcone derivatives structurally modified in propenone moiety and examined inhibitory effect on nitric oxide (NO) production and its potential mechanisms. Among the chalcone derivatives used for this study, TI-I-174 (3-(2-Hydroxyphenyl)-1-(thiophen-3-yl)prop-2-en-1-one) most potently inhibited lipopolysaccharide (LPS)-stimulated nitrite production in RAW 264.7 macrophages. TI-I-174 treatment also markedly inhibited inducible nitric oxide synthase (iNOS) expression. However, TI-I-174 did not significantly affect production of IL-6, cyclooxygenase-2 (COX-2) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), implying that TI-I-174 inhibits production of inflammatory mediators in a selective manner. Treatment of macrophages with TI-I-174 significantly inhibited transcriptional activity of activator protein-1 (AP-1) as determined by luciferase reporter gene assay, whereas nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activity was not affected by TI-I-1744. In addition, TI-I-174 significantly inhibited activation of c-Jun-N-Terminal kinase (JNK) without affecting ERK1/2 and p38MAPK, indicating that down-regulation of iNOS gene expression by TI-I-174 is mainly attributed by blockade of JNK/AP-1 activation. We also demonstrated that TI-I-174 treatment led to an increase in heme oxygenase-1 (HO-1) expression both at mRNA and protein level. Transfection of siRNA targeting HO-1 reversed TI-I-174-mediated inhibition of nitrite production. Taken together, these results indicate that TI-I-174 suppresses NO production in LPS-stimulated RAW 264.7 macrophages via induction of HO-1 and blockade of AP-1 activation.

• 대한본초학회지
• /
• 제20권2호
• /
• pp.7-16
• /
• 2005

Objectives : Scrophulariae Radix (SRE) is commonly used in combination with other herbs as a nutrient and health strengthening agent, and to remove 'heat' and replenish vital essence. The water-based extract of this herb can lower blood pressure in both anesthetized and concious animals, and exhibits an anti-inflammatory activity. But, there is lack of studies regarding the effects of SRE on the immunological activities in molecular levels. The present study was conducted to evaluate the effect of SRE on the regulatory mechanism of cytokines and nitric oxide (NO) in Raw 264.7 cells. Method : After the treatment of Scrophulariae Radix methanol extract, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. COX-2 and iNOS were determined by Immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results : Results provided evidence that SRE inhibited the production of nitrite and nitrate (NO), inducible nitric oxide synthase (iNOS), $interleukin-1{\beta}\;(IL-1{\beta})$ and interleukin-6 (IL-6), and the activation of phospholylation of inhibitor ${\kappa}B{\alpha}\;(p-I{\kappa}B{\alpha})$ in Raw 264.7 cells activated with lipopolysaccharide (LPS). Conclusion : These findings suggest that Scrophulariae Radix can produce anti-inflammatory effect, which may playa role in adjunctive therapy in Gram-negative bacterial infections.

• 동의생리병리학회지
• /
• 제21권3호
• /
• pp.721-727
• /
• 2007

Scorpion (SCP) has been clinically used for the treatment of endogenous wind to relieve convulsion, clearing away toxins, resolving hard masses and removing obstruction in the collaterals to relieve pain. Recent studies showed that scorpion toxins that affect the activating mechanism of sodium channels and indian black scorpion venom induced anti-proliferative and apoptogenic activity against human leukemic cell lines U937 and K562. There is lack of studies regarding the effects of SCP on the immunological activities. The present study was conducted to evaluate the effect of SCP on the regulatory effects of cytokines and nitric oxide (NO) for the immunological activities in Raw 264.7 cells. After the treatment of SCP MeOH extract dissolved in media for 1 h prior to the addition of lipopolysaccharide (LPS: 1 ${\mu}$g/ml), cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. Inducible nitric oxide synthase (iNOS) was determined by immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. As results, SCP inhibited the production of nitrite and nitrate (0.3 and 1.0 mg/ml), iNOS and p-$I_KB_{\alpha}$ protein, tumor necrosis factor-${\alpha}$ (0.3 and 1.0 mg/ml), interleukin-1${\beta}$ (0.3 and 1.0 mg/ml) and interleukin-6 (1.0mg/ml) in Raw 264.7 cells activated with LPS. These findings suggest that SCP can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.

• 한국발생생물학회지:발생과생식
• /
• 제1권2호
• /
• pp.165-172
• /
• 1997

the pupose of this study was to investigate the effects of gonadotropin and nitric oxide (NO) on the expression of mouse follicular bad and bax genes that are known induce apoptosis. Large and midium size follicles of immature mice were obtained at 0, 24, and 48 hours time intervals after Pregnant Mare's Serum gonadotropins(PMSG, 5 I.U.) injection. Preovulatory follicles collected at 24 hrs after PMSG injection were cultured with or without various chemicals such as gonadotropin, gonadotropin Releasing hormone(GnRH), testosterone, Sodium nitroprusside (SNP) for 24 hrs at $37^{\circ}C$. After 24 hrs culture, the culture media was used for nitrite assay and total RNA was extracted, subjected to RT-PCT for the analyses of bad and bax expression. We found that expression of bad and bax genes in follicles was markedly reduced before and after in vivo priming with hCG. When the preovulatory follicles were cultured for 24 hrs in culture media with PMSG and hCG, the expression of bad and bax genes was decreased. Moreover, SNP (NO generating agent) can significantly suppress the expression of bad and bax genes in follicles when apoptosis was induced by GnRH agonist and testosterone. At the same time, nitrite production of culture media was increased in GnRH agonist + SNP, testosterone + SNP and SNP treated groups than control group. These data demonstrated for the first time that peptide hormones and NO may play important roles in the regulation of mouse follicular differentiation and may prevent apoptosis via supressing the expression of bad and bax genes.

• Biomolecules & Therapeutics
• /
• 제21권4호
• /
• pp.258-263
• /
• 2013

We demonstrate herein that silibinin, a polyphenolic flavonoid compound isolated from milk thistle (Silybum marianum), inhibits LPS-induced activation of macrophages and production of nitric oxide (NO) in RAW 264.7 cells. Western blot analysis showed silibinin inhibits iNOS gene expression. RT-PCR showed that silibinin inhibits iNOS, TNF-${\alpha}$, and $IL1{\beta}$. We also showed that silibinin strongly inhibits p38 MAPK phosphorylation, whereas the ERK1/2 and JNK pathways are not inhibited. The p38 MAPK inhibitor abrogated the LPS-induced nitrite production, whereas the MEK-1 inhibitor did not affect the nitrite production. A molecular modeling study proposed a binding pose for silibinin targeting the ATP binding site of p38 MAPK (1OUK). Collectively, this series of experiments indicates that silibinin inhibits macrophage activation by blocking p38 MAPK signaling.

• 대한한의학회지
• /
• 제27권2호
• /
• pp.232-243
• /
• 2006

Objectives : Albizzia Julibrissin was formulated to contain various natural products known to cure erectile dysfunction. This study was aimed to investigate the effects of Albizzia Julibrissin on the nitric oxide synthase (NOS) activity, nitrite level, antioxidation and erectile responses induced by ethanol in corpus cavernosum penis of rats. Methods : The crushed Albizzia Julibrissin was extracted 3 times, each time with 3 volumes of methyl alcohol at $60^{\circ}C$ for 24 h. The extract was filtered and evaporated under a reduced pressure using a rotary evaporator to yield 45.3 g. Albizzia Julibrissin extract was oral-administered 100 mg per 1 kg of body weight for 20 days, while the normal group was administered only with a saline. The efficacy of Albizzia Julibrissin against erectile function was examined as described in the text. Results : The level of urethral NOS activity and nitrite were increased by Albizzia Julibrissin. The level of lipid peroxide was decreased by Albizzia Julibrissin. The level of urethral lipid peroxide in the ethanol-Albizzia Julibrissin double administered rats was decreased as low as in the norma! group, while the one in the ethanol-treated group was increased. The level of urethral nitrite, NOS activity, glutathione and serum testosterone in the ethanol-Albizzia Julibrissin double administered rats were as high as in the normal group, while the one in the ethanol-treated group was decreased. The erectile response to cavernous nerve stimulation in the ethanol-Albizzia Julibrissin double administered rats increased as high as in the normal group while the one in the ethanol-treated group decreased. Conclusions : Albizzia Julibrissin was shown to be effective for the treatment of erectile dysfunction induced by ethanol in rats.