• Proceedings of the Korean Society of Applied Pharmacology
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• 1998.11a
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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.

• Microbiology and Biotechnology Letters
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• v.41 no.2
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• pp.253-262
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• 2013

In this study, we analyzed the anti-oxidative and anti-inflammatory activities of seven medicinal herbs. All extracts of the tested herbs, Euryale ferox Salisbury, Echinops setifer Iljin, Amomum cardamomum Linne, Tetrapanax papyriferus, Illicium verum Hook. f., Typha orientalis Presl, and Piper longum Linne, exhibited potent anti-oxidative activity as confirmed by DPPH radical scavenging capacity. Lipopolysaccharide (LPS) induced nitric oxide (NO) production, in the RAW 264.7 cell line, was also ameliorated by all extracts' treatments in a dose dependent manner. NO suppressive activity originated from the inhibition of inducible nitric oxide synthase (iNOS) protein expression by the extracts. Three extracts, E. ferox S., I. verum Hook. f., and P. longum L., possessed suppressive activity against, not only iNOS, but also cycloxygenase 2 (COX-2) protein expression. These three extracts may then serve as potential candidates for non steroidal analgesic inflammation drugs (NSAIDs). Furthermore, all extracts induced anti-oxidative enzyme, heme oxygenase 1, protein expression. Taken together, these results provide an important new insight into the fact that various medicinal herbs possess potent anti-oxidative and anti-inflammatory activities and might be utilized as promising agents in the field of health products. Further studies for the identification of the active compounds from medicinal herbs are clearly needed.

• Journal of Applied Biological Chemistry
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• v.50 no.3
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• pp.160-163
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• 2007

Two flavonoids (1 and 2) and one phenolic acid (3) obtained from Erigeron annuus have recently been shown to have potent antioxidant activities. Aim of this study was to investigate the inhibitory effects of these components on $interferon-{\gamma}$ and lipopolysaccharide-induced nitric oxide productions in the mouse pheritoneal macrophage. Compounds 2 and 3 showed marked inhibitory activities against inducible nitric oxide synthase (iNOS) on the lipopolysaccharide and $interferon-{\gamma}-stimulated$ mouse pheritoneal macrophages without cytotoxicity. Therefore, these results suggest that the compounds could be effective anti-inflammatory agents as nitric oxide inhibitors in vivo.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.34 no.1
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• pp.28-35
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• 2008

Purpose: The nitric oxide (NO) release by inducible nitric oxide synthase (iNOS) is the key events in macrophage response to lipopolysaccharide (LPS) which is suggested to be a crucial mediator for inflammatory and innate immune responses. NO is an important mediator involved in many host defense action and may also lead to a harmful host response to bacterial infection. However, given the importance of iNOS in a variety of pathophysiological conditions, control of its expression and signaling events in response to LPS has been the subject of considerable investigation. Materials and Methods: The Raw264.7 macrophage cell line was used to observe LPS-stimulated iNOS expression. The expression of iNOS is observed by Western blot analysis and real-time RT-PCR. Protein kinase C $(PKC)-{\alpha}$ overexpressing Raw264.7 cells are established to determine the involvement of $PKC-{\alpha}$ in LPS-mediated iNOS expression. $NF-{\kappa}B$ activity is measured by $I{\kappa}B{\alpha}$ degradation and $NF-{\kappa}B$ luciferase activity assay. Results: We found that various PKC isozymes regulate LPS-induced iNOS expression at the transcriptional and translational levels. The involvement of $PKC-{\alpha}$ in LPS-mediated iNOS induction was further confirmed by increased iNOS expression in $PKC-{\alpha}$ overexpressing cells. $NF-{\kappa}B$ dependent transactivation by LPS was observed and $PKC-{\alpha}$ specific inhibitory peptide abolished this activation, indicating that $NF-{\kappa}B$ activation is dependent on $PKC-{\alpha}$. Conclusion: Our data suggests that $PKC-{\alpha}$ is involved in LPS-mediated iNOS expression and that its downstream target is $NF-{\kappa}B$. Although $PKC-{\alpha}$ is a crucial mediator in the iNOS regulation, other PKC isozymes may contribute LPS-stimulated iNOS expression. This finding is needed to be elucidated in further study.

• The Korean Journal of Physiology and Pharmacology
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• v.27 no.1
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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.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.12
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• pp.1685-1690
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• 2009

In this study, we investigated the effect of C. zawadskii extract on nitric oxide (NO) production, prostaglandin E2 (PGE2) production, protein and mRNA expression of inducible nitric oxide synthase (iNOS) in LPS-induced RAW 264.7 macrophage cells. C. zawadskii extract (5~50 μg/mL) significantly inhibited LPS-induced NO production in a concentration-dependent manner ranging from 23.3% to 100%. Consistent with the inhibitory effect on NO production, C. zawadskii extract inhibited the protein expression and mRNA expression of iNOS. Although flower extracts of C. zawadskii was not effective on the expression of PGE2 and COX-2, flower extracts of C. zawadskii, however, showed a strong anti-inflammatory activity through inhibition of NO production and iNOS expression. The present results suggest that C. zawadskii extract has an inhibitory effect on NO production, and thus can be used as an anti-inflammatory agent.

• Natural Product Sciences
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• v.18 no.3
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• pp.141-146
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• 2012

Veronica peregrina (Scrophylariaceae) has been widely used as a Korean traditional medicine for the treatment of various pathological conditions including infection, hemorrhage and gastric ulcer. In the current study, we investigated the inhibitory effect of methanolic extracts of V. Peregrina (VPM) on the LPS-mediated nitric oxide (NO) production in mouse (C57BL/6) peritoneal macrophages. NO production was significantly down-regulated by the treatment of VPM dose dependently. To evaluate the mechanism of the inhibitory action of VPM on NO production, we performed iNOS enzyme activity assay and checked the change of inducible nitric oxide synthase (iNOS) levels by Western blotting. Although VPM did not affect iNOS enzyme activity, iNOS protein expression was attenuated by VPM indicating VPM inhibits NO production via suppression of iNOS enzyme expression. In addition, VPM attenuated the expression of another pro-inflammatory mediator such as cyclooxygenase-2 (COX-2) in a dose dependent manner. We also found that VPM can reduce trypsin-induced paw edema in mice. Based on this study, we suggest that V. peregrina may be beneficial in diseases which related to macrophage-mediated inflammatory disorders.

• Biomolecules & Therapeutics
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• v.16 no.3
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• pp.231-236
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• 2008

The current work was designed to assess novel pharmacological activities of 4-hydroxybenzaldehyde (HD), a major phenolic constituent of various natural products of plant origin, such as Gastrodia elata Blume. HD exhibited a significant inhibition in the chick chorioallantoic membrane (CAM) angiogenesis. HD also displayed an inhibitory effect in acetic acid-induced permeability in mice. Anti-nociceptive activity of HD was convinced using the acetic acid-induced writhing test in mice. HD was able to suppress production of nitric oxide (NO) and induction of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in the lipopolysaccharide (LPS)-activated RAW264.7 macrophages. HD also diminished the reactive oxygen species (ROS) level elevated in the LPS-activated macrophages. In brief, HD exhibits anti-angiogenic, anti-inflammatory and anti-nociceptive activities possibly via down-regulating iNOS and/or COX-2, which may be partly responsible for pharmacological efficacies of various natural products.

• Animal cells and systems
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• v.13 no.2
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• pp.105-112
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• 2009

In this study, the effects of iron on cytochrome c oxidase (CcO) in rat lung mitochondria were examined. Similar to liver mitochondria, iron accumulated considerably in lung mitochondria (more than 2-fold). Likewise, the reactive oxygen species and nitric oxide (NO) content of mitochondria were increased by more than 50% and 100%, respectively. NO might be produced by nitric oxide synthase (NOS), eNOS and iNOS type, with particular contribution by NOS in mitochondria. The respiratory control ratio of iron overloaded lung mitochondria dropped to nearly 50% due to increased state 4. Likewise, cytochrome c oxidase activity was lowered significantly to approximately 50% due to excess iron. Real-time PCR revealed that the expression of isoforms 1 and 2 of subunit IV of CeO was enhanced greatly under excess iron conditions. Taken together, these results show that oxidative phosphorylation within lung mitochondria may be influenced by iron overload through changes in cytochrome c oxidase and NO.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.207-207
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• 1996

Through molecular cloning, five muscarinic receptors have been identified. The muscarinic receptors can be generally grouped according to their coupling to either stimulation of phospholipase C (m1, m3, and m5) or the inhibition of adenylate cyclase (m2 and m4). Each m1, m3, and m5 receptors has the additional potential to couple to the activation of phospholipase A$_2$, C, and D, tyrosine kinase, and the mobilization of Ca$\^$2+/. However, the differences in coupling efficiencies to different second messenger systems between these receptors have not been studied well. Ectopic expression of each of these receptors in mammalian cells has provided the opportunity to evaluate the signal transduction of each in some detail. In this work we compared the coupling efficiencies of the m1, m3 and m5 muscarinic receptors expressed in chinese hamster ovary (CHO) cells to the Ca$\^$2+/ mobilization and the stimulation of neuronal nitric oxide synthase (nNOS). Because G protein/PLC/PI turnover/[(Ca$\^$2+/])i/NOS pathway was supposed as a main pathway for the production of nitric oxide via muscarinic receptors, we studied on ml, m3 and m5 receptors. Stimulation of guanylate cyclase activity in detector neuroblastoma cells was used as an index of generation nitric oxide (NO) in CHO cells. The agonist carbachol increased the cGMP formation and the intracellular [Ca$\^$2+/] in concentration dependent manner in three types of receptors and the increased cGMP formation was significantly attenuated by scavenger of NO or inhibitor of NOS. m5 receptors was most efficiently coupled to stimulation of nNOS, And, the coupling efficiencies to the stimulation of neuronal nitric oxide synthase in three types of receptors were parallel with them to the Ca$\^$2+/ mobilization.