• 대한한방내과학회지
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• 제30권2호
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• pp.288-297
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• 2009

Objective : Inflammatory cytokines have a close relationship to insulin dependent diabetes mellitus (IDDM). The inhibitory effect of Smilacis Glabrae Rhizoma (SGR) were examined on production of nitric oxide (NO), prostaglandin $E_2$ $(PGE_2)$, synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and NF-${\kappa}$B activation in Raw 264.7 cells. Methods: Raw 264.7 cells were pretreated with SGR(20, 50, 100 ${\mu}g$/ml), and then cultured with lipopolysaccharides (LPS). Cell viability was measured by MTT assay; inhibition of NO, $PGE_2$, and TNF-${\alpha}$ production were measured by Griess reagent and enzyme-linked immunosorbent assay(ELISA). Induction of COX-2 and iNOS were determined by western blotting analysis. Inhibition of NF-${\kappa}$B was measured by immunofluorescence assay (IFA). Results: SGR inactivated NF-${\kappa}$B, and inhibited the production of NO, iNOS, and $PGE_2$. Inhibition of COX-2 and TNF-${\alpha}$ could not be confirmed. Conclusions: From the above result. SGR was found to have an anti-inflammatory effect of inhibition of NO, iNOS, and $PGE_2$ production via inhibition of NF-${\kappa}$B.

• The Korean Journal of Physiology and Pharmacology
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• 제9권4호
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• pp.215-221
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• 2005

We investigated the effects of testosterone and dihydrotestosterone on inflammatory response of iNOS and COX-2 expression in rat vascular smooth muscle cells. Rat vascular smooth muscle cells (VSMC) stimulated with bacterial lipopolysaccharide $(LPS;\;10{\mu}g/ml)$ for 24 hours were incubated with increasing amounts of testosterone and dihydrotestosterone (1 and 100 nM). LPS was found to induce inflammatory response of iNOS and COX-2 mRNA and protein in VSMC. These processes were affected by male sex steroid hormones. For 3 hours, however, pretreatment of the cells with 100 nM each of testosterone and dihydrotestosterone suppressed LPS induced iNOS and COX-2 protein expression. RT-PCR analysis revealed that testosterone and dihydrotestosterone did not inhibit mRNA expression of iNOS and COX-2 stimulated by 24 hours of LPS incubation. Proliferation rate was slower in VSMC treated with testosterone and dihydrotestosterone. Testosterone enhanced androgen receptor expression, and LPS significantly reduced androgen receptor protein expression in VSMC. These results indicate that the expression of both iNOS and COX-2 proteins was suppressed by testosterone and dihydrotestosterone in LPS stimulated VSMC and leading to reduction of vascular inflammation.

• The Korean Journal of Physiology and Pharmacology
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• 제8권3호
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• pp.161-165
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• 2004

Exogenous carbon monoxide (0.2%) increases L-type calcium $(Ca^{2+})$ current in human jejunal circular smooth muscle cells. The stimulatory effect of carbon monoxide (CO) on L-type $Ca^{2+}$ current is inhibited by pre-application of L-NNA, a classical competitive inhibitor of nitric oxide synthase (NOS) with no significant isoform selectivity (Lim, 2003). In the present study, we investigated which isoform of NOS affected CO induced stimulation of L-type $Ca^{2+}$ current in human jejunal circular smooth muscle cells. Cells were voltage clamped by whole-cell mode patch clamp technique, and membrane currents were recorded with 10 mM barium as the charge carrier. Before the addition of CO, cells were pretreated with each inhibitor of three NOS isoforms for 15 minutes. CO-stimulating effect on L-type $Ca^{2+}$ current was partially blocked by N-(3-(Amino-methyl) benzyl) acetamidine 2HCl (1400W, an iNOS inhibitor). On the other hand, 3-bromo-7-nitroindazole (BNI, a nNOS inhibitor) or $N^5-(1-Iminoethyl)-L-ornithine$ dihydrochloride (L-NIO, an eNOS inhibitor) completely blocked the CO effect. These data suggest that low dose of exogenous CO may stimulate all NOS isoforms to increase L-type $Ca^{2+}$ channel through nitric oxide (NO) pathway in human jejunal circular smooth muscle cells.

• BMB Reports
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• 제54권10호
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• pp.516-521
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• 2021

Although arginase primarily participates in the last reaction of the urea cycle, we have previously demonstrated that arginase II is an important cytosolic calcium regulator through spermine production in a p32-dependent manner. Here, we demonstrated that rhaponticin (RPT) is a novel medicinal-plant arginase inhibitor and investigated its mechanism of action on Ca²⁺-dependent endothelial nitric oxide synthase (eNOS) activation. RPT was uncompetitively inhibited for both arginases I and II prepared from mouse liver and kidney. It also inhibited arginase activity in both aorta and human umbilical vein endothelial cells (HUVECs). Using both microscope and FACS analyses, RPT treatments induced increases in cytosolic Ca²⁺ levels using Fluo-4 AM as a calcium indicator. Increased cytosolic Ca²⁺ elicited the phosphorylations of both CaMKII and eNOS Ser1177 in a time-dependent manner. RPT incubations also increased intracellular L-arginine (L-Arg) levels and activated the CaMKII/AMPK/Akt/eNOS signaling cascade in HUVECs. Treatment of L-Arg and ABH, arginase inhibitor, increased intracellular Ca²⁺ concentrations and activated CaMKII-dependent eNOS activation in ECs of WT mice, but, the effects were not observed in ECs of inositol triphosphate receptor type 1 knockout (IP3R1^-/-) mice. In the aortic endothelium of WT mice, RPT also augmented nitric oxide (NO) production and attenuated reactive oxygen species (ROS) generation. In a vascular tension assay using RPT-treated aortic tissue, cumulative vasorelaxant responses to acetylcholine (Ach) were enhanced, and phenylephrine (PE)-dependent vasoconstrictive responses were retarded, although sodium nitroprusside and KCl responses were not different. In this study, we present a novel mechanism for RPT, as an arginase inhibitor, to increase cytosolic Ca²⁺ concentration in a L-Arg-dependent manner and enhance endothelial function through eNOS activation.

• 대한의생명과학회지
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• 제19권3호
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• pp.180-187
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• 2013

Hypoxia is an integral part of the environment during luteolysis. In this study we examined whether hypoxia could directly stimulate endothelial cells to produce nitric oxide (NO). Endothelial cells were cultured in hypoxic (5% $O_2$) or normoxic (20% $O_2$) conditions and the levels of total NO, inducible NO and endothelial NO was measured. We found that hypoxia but not normoxia upregulated NO production. The increased NO levels correlated with increased inducible NO synthase (iNOS) expression whereas expression of endothelial NOS (eNOS) expression remained constant. Addition of the iNOS specific inhibitor 1400W to hypoxic cultures prevented NO production suggesting that hypoxia-induced NO production in endothelial cells was due mainly to upregulation of iNOS. We also found that prostaglandin $F_{2{\alpha}}$ (PGF) production was unaffected by hypoxia suggesting that upregulation of NO was not due to increased synthesis of PGF. In summary, we report that endothelial cells cultured under hypoxic conditions produce NO via the iNOS pathway. This study provides the importance of the relation between the hypoxic environment and the induction of NO by endothelial cells during regression of the corpus luteum in the ovary.

• 동의생리병리학회지
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• 제20권3호
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• pp.724-729
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• 2006

Many traditional herbal remedies exhibit several beneficial effects including anti-inflammation. The exact mechanism of the a-inflammato action of Panax notoginseng Buck F.H. Chen. however, has not been determined. In the present study, we have isolted the acting compound, emodin, from P. notoginseng and examined the effects of p. notoginseng and emodin on lipopolysaccharide (LPS)-induced nitric oxide (NO) production, and inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) gene expression in RAW264.7 macrophages. The results indicated that p. notoginseng concentration-dependently inhibited LPS-induced NO production. Furthermore, P. notoginseng inhibited the expression of LPS-induced iNOS and COX-2 proteins without an appreciable cytotoxic effect on RAW264.7 cells. Emodin also inhibited LPS-induced iNOS protein as potently as P. notoginseng. This was consistent with the findings that P. notoginseng but not emodin inhibited prostaglandin E2 synthesis induced Dy LPS.

• 동의생리병리학회지
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• 제20권5호
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• pp.1327-1333
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• 2006

Many traditional herbal remedies exhibit several beneficial effects including anti-inflammation. Panax notoginseng Buck F.H. Chen. is used as a therapeutic agent to stop haemorrhages and a tonic to promote health in Korean and Chinese medicine. The pharrnacokinetic profiles of the main P. notoginseng are still not accurately investigated. The exact mechanism of the anti-inflammatory action of P. notoginseng, however, has not been determined. In the present study, we examined the effect of P. notoginseng on lipopolysaccharide (LPS)-induced nitric oxide (NO) production, and inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) gene expression in RAW264.7 macrophages. The results indicated that P. notoginseng concentration-dependently inhibited LPS-induced NO production. Furthermore, P. notoginseng inhibited the expression of LPS-induced iNOS and COX-2 proteins without an appreciable cytotoxic effect on RAW264.7 cells. Berberine also inhibited LPS-induced iNOS protein as potently as P. notoginseng. This was consistent with the findings that P. notoginseng and also berberine inhibited prostaglandin E2 synthesis induced by LPS.

• Preventive Nutrition and Food Science
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• 제12권2호
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• pp.74-78
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• 2007

Fucoidan, that is high-molecular-weight sulfated polysaccharides extracted from brown seaweeds has been shown to elicit various biological activities. Here, we investigated the effects of fucoidan on cell proliferation and nitric oxide (NO) production in neuronal blastoma cell (SH-SY5Y). In the present study, we demonstrated that fucoidan treatment resulted in increase of cell proliferation and NO production. When cells were treated with amyloid-${\beta}$ (A${\beta}$) in the absence or presence of fucoidan, fucoidan recovered the cell viability decreased by A${\beta}$ peptides. To further determine whether nitric oxide synthase (NOS) is involved in proliferative effect of fucoidan, cells were treated with NOS inhibitors in the absence or presence of fucoidan. Selective constitutive nitric oxide synthase (cNOS) inhibitor, diphenylene iodonium chloride (DPI), caused a decrease of cell viability, whereas cell viability was increased by specific inducible nitric oxide synthase (iNOS) inhibitor, S-methylisothiourea (SMT), in the fucoidan-untreated cells. Treatment with fucoidan inhibited the cell viability decreased in DPI-exposed cells. In contrast, fucoidan had no effect on cell growth in SMT-treated cells, indicating that cNOS may not play a role in the proliferation of fucoidan-treated cells. The present data suggest that fucoidan has proliferative and neuroprotective effects and these effects may be associated with iNOS.

• 한국과학교육학회지
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• 제30권2호
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• pp.249-260
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• 2010

본 연구의 목적은 과학 영재를 대상으로 한 명시적 과학의 본성(NOS) 프로그램의 효과를 알아보고자하는 것이다. 참가자들은 6개월 동안 8개의 명시적 NOS 프로그램에 참가하였다. 연구 분석 자료로는 20명의 과학영재 학생을 대상으로 활동지, 프로그램 전후로 수행된 개방형 질문지(VNOS), 인터뷰를 사용하였다. 연구 결과 명시적 NOS 프로그램은 과학의 잠정성, 과학에 대한 사회적 문화적 영향에 대한 인식 변화에 도움이 되었다. 그러나 법칙과 이론의 구분, 과학의 경험적 측면에서는 과학영재들은 여전히 현대적 관점을 갖지 못했고 몇몇 영역에서는 모순되는 관점과 오개념을 드러냈다. 이 연구는 과학 영재 프로그램 개발에 있어서 명시적 NOS 프로그램과 과학 탐구의 상보적 관계를 고려할 때 두 활동이 동시에 제공되어야 한다는 점을 제언하고 있다.

• 과학교육연구지
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• 제44권3호
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• pp.273-288
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• 2020

본 연구에서는 2015 개정 교육과정 고등학교 통합과학 교과서에 나타난 과학의 본성(NOS) 분포를 분석하였다. 분석 대상은 2015 개정 교육과정으로 출판된 통합과학 교과서 5종 모두를 분석하였으며, 분석의 개념적 틀로는 과학적 소양 기반 4가지 영역의 과학의 본성(NOS)(Lee, 2013)을 활용하였다. 4가지 영역의 과학의 본성(NOS)은 1. 과학지식의 본성(nature of scientific knowledge), 2. 과학적 탐구의 본성(nature of scientific inquiry), 3. 과학적 사고의 본성(nature of scientific thinking), 그리고 4. 과학과 기술 및 사회의 상호작용의 본성(nature of interactions among science, technology, and society)이다. 분석은 2명의 분석자가 수행하였으며, 두 분석자간의 신뢰도는 Cohen's kappa 계수 0.83 ~ 0.96으로 비교적 높은 신뢰도 값을 나타냈다. 분석 결과는 다음과 같다. 첫째, 2015 개정 교육과정 통합과학 교과서에서는 과학의 본성(NOS) 4가지 측면 중에서 '과학탐구의 본성(nature of scientific inquiry)' 영역을 전반적으로 가장 강조하고 있었다. 이것은 통합과학 교과서 5개 출판사 전체 영역에서 '과학 탐구의 본성(범주 II)'의 분포가 평균 약 44 %로 나타나는 것에서 확인할 수 있었다. 둘째, 2015 개정 교육과정 통합과학 교과서는 출판사에 상관없이 '과학탐구의 본성(범주 II)'을 가장 강조하고 있었지만, 그외 다른 측면의 과학의 본성(NOS) 부분은 출판사별로 다소 차이가 나타나고 있었다. 따라서 통합과학 교과서들은 과학적 내용과 활동을 탐구하는 방법으로서 주로 제시하면서 출판사별로 다소 다르게 과학의 본성(NOS) 특징을 강조하고 있다고 말할 수 있다. 셋째, 2015 개정 교육과정 통합과학 교과서에서 강조하는 과학의 본성(NOS) 측면은 4. 환경과 에너지 단원을 제외하고 모든 단원에서 유사하게 나타났다. 이것은 과학의 본성(NOS) 단원별 분석 결과가 대부분의 단원에서 전체 교과서 분석 결과와 유사한 양상이었으며, 4. 환경과 에너지 단원에서만 범주 II의 '과학 탐구의 본성' 영역 다음으로 '과학-기술-사회와 상호작용하는 과학의 본성(STS)(범주 IV)'이 강조되고 있는 것에서 확인할 수 있다. 이와 같은 결과는 2015 개정 교육과정 통합 과학 교과서가 지난 교육과정에 비하여 비교적 다양하고 균형 있는 과학의 본성(NOS) 측면을 제시하고 있으며, 2015 개정 교육과정의 목표인 과학적 문제해결력과 창의력 증진을 위하여 과학적 탐구를 강조를 하고 있다는 것을 알 수 있었다.