• Journal of Periodontal and Implant Science
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• 제35권4호
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• pp.1081-1095
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• 2005

본 연구는 치주질환 주요 병인균주 중의 하나인 Porphyromonas gingivalis의 세균내독소가 마우스 대식 세포주인 RAW264.7 세포에서의 nitric oxide의 생성과 iNOS의 발현에 미치는 영향을 분석하고 그 기전을 규명하기 위해 수행되었다. Butanol추출법과 phenol-water법에 의해 P. gingivalis 381로부터 세균내독소를 추출하였으며, NO의 생성은 배양 상층액 내의 nitrite 농도를 측정하여 결정하였다. 또한, iNOS의 western blot 분석과 reverse transcription (RT)-PCR 산물의 분석을 수행하였다. P. gingivalis의 세균내독소는 부가적인 자극이 없는 상태에서도 iNOS의 발현과 NO 생성을 유발하였으며, NF- ${\kappa}B$, microtubule polymerization, protein tyrosine kinase, 그리고 protein kinase C 등이 P. gingivalis 세균내독소에 의한 NO 생성에 간여하는 것으로 여겨진다. 또한, P. gingivalis 세균내독소에 의한 NO 생성에는 L-arginine이 요구되었다. P. gingivalis 세균내독소에 의한 NO 생성은 염증성 치주질환의 발병과 진행에 있어 중요한 역할을 하는 것으로 여겨진다.

• 대한물리치료과학회지
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• 제9권4호
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• pp.45-52
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• 2002

Rats that are fed a fructose-rich diet develop hypertension, insulin resistance, and hypertriglyceridemia. To elucidate whether altered expression levels of endothelin-1 and nitric oxide synthase are related to the development of insulin-resistant hypertension, we examined the present study. Male Sprague-Dawley rats were fed a fructose-rich diet for 5 weeks. Systolic blood pressure significantly increased in fructose-fed rats. While serum free fatty acid and plasma nitrite/nitrate levels did not significantly differ between the fructose-fed and control groups, plasma insulin and serum triglyceride concentrations significantly increased in the former. Endothelin-1 mRNA expression in the aorta increased in fructose-fed rats. Neither the protein expression of constitutive nitric oxide synthase nor that of inducible nitric oxide synthase were significantly affected by fructose feeding. However, nitrite/nitrate levels in the aorta were significantly increased. These results suggest that an increase in vascular endothelin-1 is an important contributing factor to the development of hypertension in fructose-fed rats. However, the vascular nitric oxide pathway may not be causally related to the development of fructose-induced hypertension.

• The Korean Journal of Physiology and Pharmacology
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• 제5권1호
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• pp.41-46
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• 2001

To investigate the mutual relationship between angiotensin II (Ang II) and nitric oxide (NO) in paraventricular nucleus (PVN), Ang II receptor type Ia $(AT_{1A}),$ type Ib $(AT_{1B}),$ endothelial constitutive nitric oxide synthase (ecNOS), and neuronal constitutive nitric oxide synthase (ncNOS) mRNA levels of rat PVN were measured after unilateral carotid artery ligation. $AT_{1A}$ and $AT_{1B}$ mRNA levels were markedly elevated 6 hrs after unilateral carotid artery ligation. Losartan injection $(10\;{\mu}g/0.3\;{\mu}l)$ into the PVN augmented of the increment of $AT_{1A}$ and $AT_{1B}$ mRNAs It also increased ecNOS gene expression. In addition, $AT_{1B}$ mRNA levels increased after N-nitro-L-arginine methyl ester (L-NAME) injection $(50\;{\mu}g/0.3\;{\mu}l)$ into the PVN. These results suggest that Ang II and NO in the rat PVN may interplay, at least in part, through regulation of gene expression of ecNOS and $AT_{1B},$ respectively.

• 대한의생명과학회지
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• 제25권2호
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• pp.196-200
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• 2019

Effect of retinoids, i.e., all-trans retinoic acid and 13-cis retinoic acid, on the activity of nitric oxide synthase (NOS) was evaluated in human malignant keratinocytes to examine the possible correlation of retinoids with NOS activities. All-trans retinoic acid and 13-cis retinoic acid did not alter the nitric oxide (NO) production. However, in the presence of lipopolysaccharide (LPS, $1{\mu}g/mL$), they significantly increased NO release in a dose-dependent manner until 48 h at concentrations of $50{\sim}100{\mu}M$. The degree of upregulation of NO by all-trans retinoic acid and 13-cis retinoic acid increased up to 35% and 37%, respectively, compared to that by the control, which demonstrated the upregulation of LPS-inducible nitric oxide synthase (iNOS)-dependent generation of NO as well as showing a crucial link between retinoids-induced activity and NOS. Findings of this study now suggest that the upregulation of LPS-iNOS activity may be associated with modulation of retinoids-induced control of cellular developmental processes, which may produce new therapeutics of retinoids in the complexity of how NO affects human keratinocytes.

• 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.141-141
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• 1998

In order to study the effect of nitric oxide on the regulation of mouse cyplal expression, 5' flanking DNA of mouse cytochrome P450 lal was cloned into pGL3 basic vector encoding luciferase gene. pcyplal-Luc was transfected into Hepa I cells and various chemicals were treated. Luciferase activity was stimulated 1000 folds over that of control by TCDD (2,3,7,8-tetrachloro-p-dioxin) treatment and this stimulation was dose dependent. When SNP (sodium nitroprusside) which donates nitric oxide was administrated, this stimulatory effect of TCDD on luciferase activity was decreased. And LPS (lipopolysaccharide) which is an iNOS (inducible nitric oxide synthase) inducer also decreased the stimulatory effect of TCDD on luciferase. And iNOS inhibitor N$\^$G/-nitro-ι-arginine + TCDD treatment increased the stimulation effect of TCDD and this effect was abolished when ι-arginine was added to N$\^$G/-nitro-ι-arginine + TCDD treatment. When N$\^$G/-nitro-ι-arginine was concomitantly administrated with SNP or LPS to confirm the effect of nitric oxide, the inhibitory effect of SNP or LPS was abolished. These data strongly suggest that nitric oxide might be an inhibitory regulator on the cytochrome P450 lal gene expression in Hepa cells.

• 생명과학회지
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• 제23권6호
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• pp.743-750
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• 2013

본 연구의 목적은 혈관계 순환을 촉진시키는 약효제를 발굴하는 것이다. 아사이베리(Acai berry) 에탄올 추출물(ABEE)의 항산화 효과 뿐만 아니라 토끼의 폐에서 유래한 angiotensin converting enzyme (ACE)의 활성 및 대식세포와 혈관내피세포에서 nitric oxide 생성에 대한 효과가 본 연구에서 조사되었다. 먼저 환원력과 지질과산화에 대한 ABEE의 항산화효과가 관찰되었다. 뿐만 아니라, ABEE는 hydroxyl radical에 의하여 유발된 DNA 산화에 대한 보호효과를 보여주었다. 더욱이 ABEE는 0.01%의 농도에서 약 50% 정도의 angiotensin converting enzyme 활성에 대한 억제효과를 발휘하는 것으로 나타났다. 혈관내피세포에서 ABEE는 nitric oxide 의 생성을 증가시켰으나 대식세포에서는 lipopolysaccharide에 의하여 생성된 nitric oxide 생성을 감소시켰다. 게다가 superoxide dismutase (SOD)-2와 -3 발현수준은 ABEE 처리에 의하여 증가되었으나 SOD-1의 발현수준은 일정하였다. 더욱이 nitric oxide synthases-1 (NOS-1)의 발현수준은 ABEE 처리에 의하여 증가되었으나, 유발 효소인 NOS-2의 발현수준은 일정하였다. 또한 SOD의 전사인자인 Nrf-2의 발현수준은 ABEE에 의하여 증가되었다. 그러므로 이러한 결과들은 ABEE가 이상의 작용을 경유하여 혈액순환을 촉진시킬 수 있고, 혈관의 건강을 위한 큰 잠재성을 가지고 있다는 것을 암시하고 있다.

• The Korean Journal of Physiology and Pharmacology
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• 제27권1호
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• pp.9-20
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• 2023

The mechanism is unclear for the reported protective effect of hyperbaric oxygen preconditioning against oxidative stress in tissues, and the distinct effects of hyperbaric oxygen applied after stress. The trained mice were divided into three groups: the control, hyperbaric oxygenation preconditioning, and hyperbaric oxygenation applied after mild (fasting) or hard (prolonged exercise) stress. After preconditioning, we observed a decrease in basal levels of nitric oxide, tetrahydrobiopterin, and catalase despite the drastic increase in inducible and endothelial nitric oxide synthases. Moreover, the basal levels of glutathione, related enzymes, and nitrosative stress only increased in the preconditioning group. The control and preconditioning groups showed a similar mild stress response of the endothelial and neuronal nitric oxide synthases. At the same time, the activity of all nitric oxide synthase, glutathione (GSH) in muscle, declined in the experimental groups but increased in control during hard stress. The results suggested that hyperbaric oxygen preconditioning provoked uncoupling of nitric oxide synthases and the elevated levels of GSH in muscle during this study, while hyperbaric oxygen applied after stress showed a lower level of GSH but higher recovery post-exercise levels in the majority of antioxidant enzymes. We discuss the possible mechanisms of the redox response and the role of the nitric oxide in this process.

• 대한약침학회지
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• 제12권4호
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• pp.89-96
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• 2009

목적:이 연구의 목적은 유산균발효애엽 물추출물이 에탄올 등으로 약화된 마우스 대식세포의 NO 생성에 미치는 영향을 조사하는 것이다. 방법:애엽을 유산균으로 발효시켜 시료(AFL)를 만들고 만들어진 시료를 10, 50, 100, 200, 400 ug/mL의 농도로 에탄올, 갈릭산, 아세트아미노펜, 아세트알데히드, 니코틴과 함께 24시간동안 마우스 대식세포에 처리한 후 세포배양액을 채취, NO 생성정도를 측정하여 비교하였다. 결과:1. AFL은 400 ug/mL의 농도에서 갈릭산에 의한 마우스 대식세포의 NO생성억제를 유의하게 회복시켰다. 2. AFL은 200, 400 ug/mL의 농도에서 에탄올에 의한 마우스 대식세포의 NO생성억제를 유의하게 회복시켰다. 3. AFL은 400 ug/mL의 농도에서 니코틴에 의한 마우스 대식세포의 NO생성억제를 유의하게 회복시켰다. 4. AFL은 200, 400 ug/mL의 농도에서 아세트아미노펜에 의한 마우스 대식세포의 NO생성억제를 유의하게 회복시켰다. 5. AFL은 200, 400 ug/mL의 농도에서 아세트알데히드에 의한 마우스 대식세포의 NO생성억제를 유의하게 회복시켰다. 결론:유산균발효애엽추출물(AFL)은 에탄올, 갈릭산, 니코틴, 아세트알데히드, 아세트아미노펜 등에 의해 약화된 대식세포의 NO생성을 회복시킴으로서 다양한 독성물질에 의하여 약화되는 식세포의 항병능력을 회복시키는 면역강화물질로 개발될 수 있을 것이다.

• Journal of Trauma and Injury
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• 제21권1호
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• pp.59-65
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• 2008

Purpose: The aim of this study was to evaluate the effect of overexpression of heat shock protein 70 (HSP70) on the expression of inducible nitric oxide synthase and on the concentration of nitric oxide and to determine the mechanism for the relationship between HSP70 and inducible nitric oxide synthase (iNOS) in sepsis. Methods: Experiments were performed on male Sprague-Dawley rats, and sepsis was induced by using cecal ligation and puncture (CLP). Glutamine (GLN) or saline was administered 1 h after initiation of sepsis. We acquired serum and lung tissues from the rats 12 h or 24 h after initiation of sepsis. We analyzed the concentration of nitric oxide, the expression of HSP70 in the lung, and the gene expression of iNOS in the lung. Results: In CLP+GLN, glutamine given after initiation of sepsis enhanced the expression of HSP70 in the lung at 12 h (CLP+GLN vs. CLP:: $47.19{\pm}10.04$ vs. $33.22{\pm}8.28$, p = 0.025) and 24 h (CLP+GLN vs. CLP: $47.06{\pm}10.60$ vs. $31.90{\pm}4.83$, p = 0.004). In CLP+GLN, glutamine attenuated the expression of iNOS mRNA in the lung at 12 h (CLP+GLN vs. CLP: $4167.17{\pm}951.59$ vs. $5513.73{\pm}1051.60$, p = 0.025) and 24 h (CLP+GLN vs. CLP: $9,437.65{\pm}2,521.07$ vs. $18,740.27{\pm}8,241.20$, p = 0.016) and reduced the concentration of nitric oxide in serum at 12 h (CLP+GLN vs. CLP: $0.86{\pm}0.48$ vs. $3.82{\pm}2.53{\mu}mol/L$, p = 0.016) and 24 h (CLP+GLN vs. CLP: $0.39{\pm}0.25$ vs. $1.85{\pm}1.70{\mu}mol/L$, p = 0.025). Conclusion: The overexpression of HSP70 induced by the administration of glutamine in sepsis attenuated the gene expression of iNOS and reduced the concentration of nitric oxide.

• 생명과학회지
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• 제5권1호
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• pp.20-25
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• 1995

기생충감영시 cytokine으로 활성화된 대식세포가 방어기전의 Effector cell로 작용할 때 분비하는 nitric oxide의 양 및 TNF-$\alpha$ 와 IFN-$\gamma$의 분비정도와 nitric oxide와의 상관관계 등을 알아보기 위하여 3종의 흡충류, Fasciola, paragonimus, Schistosma의 조항원 (100mg/ml)을 마우스 복강내에 주사　 후 24시간, 72시간, 9일간격으로 마우스의 대식세포(1X10$^{6}$/ml)를 분리하여 RPMI 배지 (10% FCS 첨가)에서 48시간 배양후 Nitric TNF-$\alpha$ 및 IFN-$\gamma$를 ELISA로 정량하여 다음과 같은 결과를 얻었다. Nitrite 생성정도는 Fasciola 조항원으로 24시간 감작시킨 대식세포에서 가장 높게 나타났으며 (140$\mu$M/ml) Paragonimus 항원군에서는 24시간에 최고치에 달하였다가(34 $\mu$M/ml) 시간이 경과함에 따라 점차 감소하였다. IFN-$\gamma$는 Paragonimus 항원군에서만 대조군에 비해 높았으며 9일경에 최고치를 보였다(475ng/ml). TNF-$\alpha$는 Schistosoma 항원군에서는 nitric oxide의 생성과 분비 양상이 일치하였다. 위의 결과에 의하면, 흡충류항원으로 감작된 마우스 대식세포의 nitric oxide 생성에 영향을 미치는 cytokine의 종류는 흥충류에 따라 차이를 보였으며, 이 중 Paragonimus 항원에 의해서는 IFN-$\gamma$의 분비가 촉진되는 것으로 나타났고, Schistosoma 의 경우에는 TNF-$\alpha$가 nitric oxide의 생성에 관계함을 알 수 있었다.