• BMB Reports
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• v.39 no.6
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• pp.759-765
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• 2006

In this study, we investigate Nitric oxide synthase (NOS) expressions in adriamycin (ADR)-induced cadiomyopathy in rats. Sixty male Wistar rats were randomly divided into two main groups: control and ADR groups. Myocardial histopathological observation was performed; Expressions of 3 isoforms of NOS genes were examined by RT-PCR analysis; Expressions of 3 isoforms of NOS protein was assessed by Western blot analysis. Myocardium exhibited intensive morphological changes after 8 weeks of ADR treatment. The expression levels of inducible NOS (iNOS) gene and protein were significantly increased in ADR-treated rats after 8 weeks of treatment and then slightly increased at weeks 9 and 10. No significantly difference of neuronal NOS (nNOS) or endothelial NOS (eNOS) gene and protein were observed in the myocardium obtained from the control rats and ADR-injected rats at any time point. iNOS gene expression is selectively induced by ADR in heart. The upregulation of iNOS gene and protein may be somehow correlated with morphological changes seen in heart of rat treated with ADR.

• Journal of Life Science
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• v.13 no.4
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• pp.441-447
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• 2003

To examine the role of protein kinase C (PKC) in regulation of interleukin-1 beta (IL-1$\beta$)-induced iNOS expression, IL-1$\beta$-induced nitrite production was observed in cultured vascular smooth muscle (VSM) cells pretreated with phorbol 12-myristate 13-acetate (PMA) and phorbol 12,13-butyrate (PDB) as PKC activator; 4$\alpha$-phorbol-didecanoate (PDD) as PKC non-activator. Nitrite production induced by IL-1$\beta$ was increased by the presence of increasing concentration of PMA ranging from 2 to 200 nM. However, in VSM cells pretreated with PMA and PDB, IL-1$\beta$-induced $NO_2$ production was decreased in proportion to the duration of pretreatment, and most significantly decreased in pretreatment time of 24 hours. Using RT-PCR method, the expression of iNOS mRNA induced by IL-1$\beta$ was decreased in VSM cells pretreated with PMA 200 nM for 24 hours. These results suggest that decrease in IL-I$\beta$-induced nitrite production by the pretreatment of PMA result from inhibition of iNOS expression and the inhibition related to PMA-induced PKC down-regulation.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.2
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• pp.99-104
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• 2000

We investigated the action of NOS inhibitors on NOS in rats. Both of nitric oxide synthase inhibitors, $N^G$-monomethyl-L-arginine $(L-NMMA,\;3\;{\mu}M)$ or $N^G$-nitro-L-arginine methylester $(L-NAME,\;30\;{\mu}M),$ augmented phenylephrine $(PE,\;10^{-7}\;M)-induced$ contraction which was inhibited by acetylcholine (ACh) in rat thoracic aorta. This augmentation by L-NAME or L-NMMA was attenuated with the treatment of NO precursor, arginine. ACh, however, decreased the augmentation induced by L-NMMA, but not by L-NAME. Superoxide dismutase (SOD, 50 u/ml) potentiated an inhibitory effect of ACh on the PE $(10^{-7}\;M)-induced$ contraction. It has been known that platelet activating factor itself induces iNOS. Platelet activating factor $(PAF,\;10^{-7}\;M)$ inhibited PE $(10^{-7}\;M)-induced$ contraction. Pretreatment with L-NMMA (30 mM) or L-NAME (30 mM) significantly blocked the inhibitory action of PAF on PE-induced contraction. L-NMMA (100 mM) or L-NAME (100 mM) reduced nerve conduction velocity (NCV) relevant to nNOS in rat sciatic nerve. ACh attenuated the reduction of NCV by L-NMMA-, but not by L-NAME-induced reduction of NCV. These results suggest that L-NMMA and/or L-NAME have different action on three types of NOS in rats.

• Korean Journal of Acupuncture
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• v.22 no.4
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• pp.109-116
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• 2005

목적 : 골관절염은 진통을 수반하는 퇴행성 관절질환이며, 장애를 일으키는 주요한 원인이 된다. 또한 노인들에 있어서 골관절염은 매우 흔한 질환이라 할 수 있다. Nitric oxide(NO)는 Nitric Oxide Synthase(NOS)에 의하여 칼슘의존성통로를 통하여 L-arginine 으로부터 합성되어지며, NO는 중추신경계에 있어서 중요한 세포사이의 전달자이다. 방법 :본 연구에서는 위령선 으로부터 추출한 액이 콜라겐으로 유도된 관절염에 걸린 쥐의 dorsolateral periaqueductal gray(DL-PAG) 영역에서 nNOS(neuronal NOS)와 NOS에 대하여 미치는 영향 을 nNOS immunohistochemistry와 nicotinamide adenine dinucleotide phosphate-diaphorase(NADPH-d) 검사법을 통하여 조사하였다. 결과 : 골관절염이 유발된 흰쥐의 DL-PAG 영역에서 nNOS와 NOS의 발현억제가 관찰되었으며, 위령선이 콜라겐으로 유도된 골관절염에서 감소된 nNOS와 NOS의 발현이 증가되었다. 결론 : 본 연구를 통하여 위령선은 골관절염이 유발된 흰쥐의 DL-PAG에서의 nNOS와 NOS의 발현에 영향을 미친다는 결과를 얻을 수 있었다.

• The Korean Journal of Physiology and Pharmacology
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• v.10 no.6
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• pp.337-341
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• 2006

The present study was designed to investigate the protein expression of nitric oxide synthase (NOS) and guanylyl cyclase (GC) activity in ischemia/perfusion (I/R) renal injury in rats. Renal I/R injury was experimentally induced by clamping the both renal pedicle for 40 min in Sprague-Dawley male rats. The renal expression of NOS isoforms was determined by Western blot analysis, and the activity of guanylyl cyclase was determined by the amount of guanosine 3', 5'-cyclic monophosphate (cGMP) formed in response to sodium nitroprusside (SNP), NO donor. I/R injury resulted in renal failure associated with decreased urine osmolality. The expression of inducible NOS (iNOS) was increased in I/R injury rats compared with controls, while endothelial NOS (eNOS) and neuronal NOS (nNOS) expression was decreased. The urinary excretion of NO metabolites was decreased in I/R injury rats. The cGMP production provoked by SNP was decreased in the papilla, but not in glomerulus. These results indicate an altered regulation of NOS expression and guanylyl cyclase activity in I/R-induced nephropathy.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.208-208
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• 1996

In this work we investigated coupling of the m2 and m4 subtypes of muscarinic acetylcholine receptors expressed in chinese hamster ovary (CHO) cells to activation of neuronal nitric oxide synthase (nNOS). Stimulation of guanylate cyclase activity in detector neuroblastoma cells was used as an index of generation of nitric oxide (NO) in CHO cells. The agonist carbachol induced marked time and concentration-dependent enhancement of the activity of nNOS at m2 receptors. In sharp contrast, the response in CHO cells transfected with the m4 receptor gene was similar in magnitude to that observed in non-transfected cells, suggesting lack of significant coupling of m4 muscarinic receptors to NO signaling. This novel observation of functional divergence of the two muscarinic receptor subtypes at the level of activation of nNOS is quite intriguing, in light of the currently accepted dogma that they belong to the same functional class. This functional selectivity was not due to differential effects on intracellular Ca$\^$2+/ concentration, since activation of both subtypes of muscarinic receptors produced a comparable, albeit quite small, Ca$\^$2+/ signal. Taken together, our present data strongly suggest that the generally assumed functional equivalence of m2 and m4 muscarinic receptors should be carefully reexamined. These data also suggest the presence of alternate mechanisms of activation of nNOS, which might be operative in the absence of large changes in the concentration of cellular Ca$\^$2+/. The latter mechanisms are expected to be activated by m2, but not m4 muscarinic receptors. Both sets of findings are quits important in regards to refining the functional classification of muscarinic receptor subtypes and the cellular mechanisms of activation of NOS.

• Korean Journal of Acupuncture
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• v.29 no.2
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• pp.300-314
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• 2012

Objectives : This study is to observe the changes in the expression of neurotransmitters, such as NO, nNOS, and NE, upon the needle insertion to the sea points, which is one of the five transport points. Methods : Needles were inserted into rats, on both left and right sides of all sea points, including the LU5, PC3, HT3, LI11, TE10, SI8, SP9, LR8, KI10, ST36, GB34, and BL40, which are the sea points of five transport points for 12 meridian vessels. After insertion, needles were retained for five minutes. After the retention, blood was drawn via cardiac puncture, and tissues of each point near meridian vessels were extracted to examine the changes in the expression of NO, nNOS and NE. Results : In terms of the effect in NO production, there was a significant decrease only in the LU5 point, whereas there was a significant increase in the TE10 point alone. In terms of the expression of nNOS within tissues, none of the experimental groups showed significant changes based on the results of immunohistochemistry and western blotting. Regarding the formation of norepinephrine within tissues, the HT3, SP9, and KI10 point showed a significant decrease, while the PC3 and LR8 point showed a significant increase. Production of plasma norepinephrine was significantly increased at the TE10, SP9, LR8, GB34, and BL40 point. Conclusions : The effect of needles applied at the sea points of five transport points of 12 meridian vessels on the functions of NO, nNOS, and NE could be observed, and it is considered that the effect of needle stimulation on nervous system disorders could be studied through additional researches based on this one.

• Korean Journal of Acupuncture
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• v.32 no.4
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• pp.160-168
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• 2015

Objectives : This study was intended to observe the changes in the expression of neurotransmitters, such as nNOS, NO and NE upon the needle insertion at varying depths at the connecting point. Methods : Needles were inserted into rats, on both left and right sides of the connecting point, including the LI6, SI7 and TE5 acupoints which are three yang meridians of the hand. After insertion, needles were retained for five minutes. Each acupuncture groups were treated acupuncture at each acupoint and at the depths of superficial, middle and deep layer. After the retention, blood was drawn via cardiac puncture, and tissues of each point near meridian vessels were extracted to examine the changes in the expression of nNOS, NO and NE. Results : In terms of the effect in nNO production, there was a significant increase only in the middle and deep layer at SI7 acupoint, but there was no significant change in the expression of NO. Regarding the formation of norepinephrine within tissues, the middle layer on LI6 acupoint, the middle layer and the deep layer on TE5 acupoint showed a significant increase, while production of plasma norepinephrine was significantly decreased at the middle layer and the deep layer on LI6 acupoint and the deep layer on SI7 acupoint. Conclusions : The effect of needles applied at the connecting point of three yang meridians on the activities of nNOS, and NE could be observed, and it can be induced that the effect of needle stimulation on disrupted nervous system can be examined through additional researches based on this one.

• Molecules and Cells
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• v.19 no.3
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• pp.408-417
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• 2005

Nitric oxide (NO) occurs in various types of cells in the central nervous system. We studied the distribution and morphology of neuronal nitric oxide synthase (NOS)-containing neurons in the visual cortex of mouse and rabbit with antibody immunocytochemistry. We also compared this labeling to that of calbindin D28K, calretinin, and parvalbumin. Staining for NOS was seen both in the specific layers and in selective cell types. The densest concentration of intense anti-NOS immunoreactive (IR) neurons was found in layer VI, while the weak anti-NOS-IR neurons were found in layer II/III in both animals. The NOS-IR neurons varied in morphology. The large majority of NOS-IR neurons were round or oval cells with many dendrites coursing in all directions. Two-color immunofluorescence revealed that only 16.7% of the NOS-IR cells were double-labeled with calbindin D28K in the mouse visual cortex, while more than half (51.7%) of the NOS-IR cells were double-labeled with calretinin and 25.0% of the NOS-IR cells were double-labeled with parvalbumin in mouse. By contrast, 92.4% of the NOS-IR neurons expressed calbindin D28K while only 2.5% of the NOS-IR neurons expressed calretinin in the rabbit visual cortex. In contrast with the mouse, none of the NOS-IR cells in the rabbit visual cortex were double-labeled with parvalbumin. The results indicate that neurons in the visual cortex of both animals express NOS in specific layers and cell types, which do not correlate with the expression of calbindin D28K, calretinin or parvalbumin between the two animals.

• Journal of Life Science
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• v.12 no.5
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• pp.505-514
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• 2002

The aim of this study was to evaluate the anticonvulsant activity of four agents derived from tropinone (T-1: 2,4-dipyrrolylmethenylnortropinone, T-2: 2,4-diphenylmethenylnortropinone, T-3 : 2,4-difurfurylmethenylnortropinone, T-4 : 2,4-dimethoxyphenylmethenylnortropinone) in NIH Swiss mouse. Pentylenetetrazole (nZ) was injected via intraperi-toniurn in mouse and Maximal Electroshock (MES) stimulation was through both conjunctivas by electrodes. Tropinone derivatives were treated at 15 minutes before PTZ or MES procedure. PIZ of 25 mg/kg induced generalized seizure in mouse, effects of tropinone derivatives on PTZ-induced seizure were monitored. Compared with control group, T-4 decreased seizure grade most effectively. Also T-4 increased onset time of PTZ-induced seizure. This result showed that T-4 is most effective on PTZ-induced seizure. In MES-induced seizure, T-1 decreased seizure grade and recovery time. nNOS expression in hippocampus and cortex were increased in nZ- and MES-induced seizure animals compared with control. Pretreatment of tropinone derivatives in PTZ-induced seizure did not affected nNOS expression in brain tissues, but T-1 and T-4 decreased nNOS expression in cortex of MES-induced seizure animals. These findings suggest that tropinone derivatives have specific anticonvulsant activities according to PTZ- and MES-induced seizure. 2,4- dimethoxyphenylmethenylnoroopinone is most effective in PTZ-induced seizure and 2,4-di methoxyphenylmethenylnortropinone is most effective in MES-induced seizure.