• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1993년도 제2회 신약개발 연구발표회 초록집
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• pp.156-156
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• 1993

본 연구에서는 생체 뇌 국소부위의 세포외액을 대상으로 실험할 수 있는 뇌 미세투석법을 이용하여 후시상하부에서 유리되는 monoamine의 세포외액 농도에 대한 전신 혈압변화의 영향을 관찰함으로써, 심혈관 조절에 대한 후시상하부의 monoamine성 뉴론의 생리적 역할을 규명하고자 하였다. 마취한 흰쥐의 머리를 뇌정위 고정장치에 고정시키고 미세투석관 (microdialysis probe)을 후시상하부에 위치시킨 후 링거액으로 관류하였다. 미세투석액내에 존재하는 monoamine성 신경 전달물질들과 그 대사체들의 정량분석은 고속액체크로마토그라피와 전기화학검출기를 이용하여 실시하였다. 전신 혈압을 40분 동안 약 40mmHg 상승 혹은 30mmHg 감소시키기 위하여 L-phenylephrine hydrochloride (800ng/100 g/min) 혹은 nitroprusside dihydrate (500 ng/100 g/min)를 대퇴정맥을 통하여 주사하였다. 후시상하부로 부터 20분 간격으로 얻은 투석액에서 신정화학물질들의 농도는 미세투석관 삽입후 2시간에 안정되었다. 관류액의 $K^{＋}$ 농도를 90 mM로 증가시켰을 때 후시상하부의 투석액에서 norepinephrine (NE) 과 serotonin (5-HT)의 농도는 각각 기준치의 176.5 $\pm$ 14.8%, 149.1 $\pm$ 2.3%로 증가하였다. Phenylephrine (i.v.)으로 유발된 전신 혈압상승에 의하여 NE과 5-HT의 양은 각각 기준치의 79.3 $\pm$ 4.4%, 61.4 $\pm$ 10.3%로 유의하게 감소하였다. 또한 nitroprusside (i.v.)에 의하여 전신혈압이 감소하였을 때 투석액내 5-HT의 양은 기준치의 195.0 $\pm$ 23.0% 로 유의하게 증가하였다. 후시상하부의 투석액내 monoamine성 대사체들의 경우에는 전신 혈압의 변화에 대한 유의한 반응을 나타내지 않았다. 이상을 종합하면 전신 혈압이 증가되었을 때는 말초 압수용체의 흥분에서 기시한 신경충동이 후시상하부에 전달되어 신경말단에서 5-HT 과 NE 의 유리가 감소되고, 5-HT의 경우에는 전신 혈압이 감소되었을때 그 반대 현상이 일어난다는 가설을 제시할 수 있다. 본 연구의 결과는 생체상태에서 혈압변화에 대한 후시상하부의 monoamine성 조절양상을 규명한 것으로서 특히 후시상하부의 serotonin성 신경계를 통한 심혈관 중추조절의 가능성을 처음으로 제시한 것이다.

• KSBB Journal
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• 제28권4호
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• pp.260-268
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• 2013

Transgenic plant cell cultures are an attractive expression system for the production of industrial and pharmaceutical proteins because of their advantages in safety and low production cost. Human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced and secreted when sugar was depleted in culture medium by transgenic rice cell lines (Oryza sativa L.) using RAmy3D promoter. Due to the production of the target protein by sugar depletion, concomitant occurrence of cell death is inevitable. For that reason, inhibition of cell death for enhancing productivity was necessary for the production period without energy sources. Supplementation of 0.1 mM sodium nitroprusside improved cell viability by 1.4-fold and maximum hCTLA4Ig production by 1.3-fold compared to those of control. Addition of 1 and 10 mM glutathione, N-acetylcysteine (NAC), and nicotinamide inhibited apoptotic-like programmed cell death by decreasing the activity of reactive oxygen species. Production hCTLA4Ig was enhanced 1.4-, 1.25-, and 1.15-fold with 10 mM NAC, 1 mM NAC, and 1 mM glutathione, respectively. In addition, it was found that the supplementation of NAC enhanced the cell viability.

• 약학회지
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• 제45권1호
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• pp.116-124
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• 2001

Nitric oxide is a labile, gaseous, broad spectrum second messenger that used in various tissues and cells. If it is induced by endogenously and exogenously in the neuronal cells, it is able to mediate analgesia or hyperalgesia at the periphery and in the spinal level respectively. This dual role of nitric oxide in the sensory system is very intriguing but has not been fully understood yet. In this experiment, acetylcholine (300 $\mu$g/paw), sodium nitroprusside (600 $\mu$g/paw), and L-arginine (300 $\mu$g/paw) represented antinociceptive effect to noxious topical stimulus, but pronociceptive responses followed by spinally application (20$\mu$g/5$\mu$l, 10$\mu$g/3$\mu$l, 500$\mu$g/5$\mu$l respectively). Calcium ion is critical element which activates nitric oxide synthase, therefore verapamil (300 $\mu$g/paw) and NOS inhibitor (20 mg/kg, L-NAME or L-NOArg) are injected into right hind paw (i.pl.). When verapamil is combined with NOS inhibitors analgesic effects through NO-cGMP pathway are inhibited as compared with ACh alone. Diluted formalin (2.5%), when injected into rats'hind paw (0.05 ml), elicited a biphasic algesic responses and nitric oxide had an analgesic effect on both $A\delta$ and C sensory nerve fibers which manipulate the phases respective1y. Nitric oxides, which produced from constitutive nitric oxide synthase, activated cyclooxygenase-type I and then prostaglandins are produced from them. So, indomethacin and ibuprofen, inhibitors of COX$_1$enzyme, when pretreated intraperitoneally (100 mg/kg) could reduce the hyperalgesic state. From these results, it is possible to imagine that the intrathecally administered NO donors expressed hyperalgesia through both long-term potentiation mechanism and arachidonic acid-prostaglandin cascade.

• The Korean Journal of Physiology and Pharmacology
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• 제19권3호
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• pp.275-281
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• 2015

Orthostatic hypotension is most common in elderly people, and its prevalence increases with age. Attenuation of the vestibulo-sympathetic reflex (VSR) is commonly associated with orthostatic hypotension. In this study, we investigated the role of glutamate on the vestibulo-solitary projection of the VSR pathway to clarify the pathophysiology of orthostatic hypotension. Blood pressure and expression of both pERK and c-Fos protein were evaluated in the nucleus tractus solitarius (NTS) after microinjection of glutamate into the medial vestibular nucleus (MVN) in conscious rats with sodium nitroprusside (SNP)-induced hypotension that received baroreceptor unloading via sinoaortic denervation (SAD). SNP-induced hypotension increased the expression of both pERK and c-Fos protein in the NTS, which was abolished by pretreatment with glutamate receptor antagonists (MK801 or CNQX) in the MVN. Microinjection of glutamate receptor agonists (NMDA or AMPA) into the MVN increased the expression of both pERK and c-Fos protein in the NTS without causing changes in blood pressure. These results indicate that both NMDA and AMPA receptors play a significant role in the vestibulo-solitary projection of the VSR pathway for maintaining blood pressure, and that glutamatergic transmission in this projection might play a key role in the pathophysiology of orthostatic hypotension.

• Plant Biotechnology Reports
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• 제5권4호
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• pp.353-365
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• 2011

The present study investigates the possible regulatory role of exogenous nitric oxide (NO) in antioxidant defense and methylglyoxal (MG) detoxification systems of wheat seedlings exposed to salt stress (150 and 300 mM NaCl, 4 days). Seedlings were pre-treated for 24 h with 1 mM sodium nitroprusside, a NO donor, and then subjected to salt stress. The ascorbate (AsA) content decreased significantly with increased salt stress. The amount of reduced glutathione (GSH) and glutathione disulfide (GSSG) and the GSH/GSSG ratio increased with an increase in the level of salt stress. The glutathione S-transferase (GST) activity increased significantly with severe salt stress (300 mM). The ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), catalase (CAT) and glutathione peroxidase (GPX) activities did not show significant changes in response to salt stress. The glutathione reductase (GR), glyoxalase I (Gly I), and glyoxalase II (Gly II) activities decreased upon the imposition of salt stress, especially at 300 mM NaCl, with a concomitant increase in the $H_2O_2$ and lipid peroxidation levels. Exogenous NO pretreatment of the seedlings had little influence on the nonenzymatic and enzymatic components compared to the seedlings of the untreated control. Further investigation revealed that NO pre-treatment had a synergistic effect; that is, the pre-treatment increased the AsA and GSH content and the GSH/GSSG ratio, as well as the activities of MDHAR, DHAR, GR, GST, GPX, Gly I, and Gly II in most of the seedlings subjected to salt stress. These results suggest that the exogenous application of NO rendered the plants more tolerant to salinity-induced oxidative damage by enhancing their antioxidant defense and MG detoxification systems.

• Journal of Korean Neurosurgical Society
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• 제38권3호
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• pp.221-226
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• 2005

Objective : Reactive oxygen metabolites and polymorphonuclear leukocytes have been implicated in the pathophysiology of reperfusion injury. The mechanisms involved in superoxide-mediated leukocyte adherence remain unclear, however, nitric oxide[NO] may contribute to this response. The present study is undertaken to elucidate mechamisms controlling NO based mechanisms that regulated leukocyte-endothelial interactions in the cerebral vasculature after global cerebral ischemia and reperfusion. Methods : Pial venular leukocyte adherence of anesthetized newborn piglets was quantified by in situ fluorescence videomicroscopy through closed cranial windows during basal conditions and during 2hours of reperfusion after global ischemia induced by 9minutes of asphyxia. Nitric oxide synthase[NOS] was inhibited by local window superfusion of L-nitroarginine[NA]; superfusion of sodium nitroprusside[SNP] was used to donate NO. Results : The mean number of adherent leukocytes to cerebral venules in the 9minutes asphyxia and 2hours reperfusion group were $161{\pm}19$ compared with $13{\pm}4$ in the nonasphyxial group. Superfusion of L-NA through the cranial window for 2hours resulted in leukocyte adherence similar to that observed during the initial 2hours of reperfusion after asphyxia. Leukocyte adherence was not additionally increased in asphyxic animal treated with L-NA. SNP inhibited asphyxia induced leukocyte adherence back to control levels. Conclusions : Nitric oxide inhibits leukocyte adherence to cerebral venules during the initial hours of reperfusion after asphyxia, and that NO supplementation inhibit asphyxia induced leukocyte adherence back to control levels. These results indicate that NO is an important factor in ischemia-reperfusion induced leukocyte adherence.

• Reproductive and Developmental Biology
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• 제28권2호
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• pp.83-87
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• 2004

본 연구는 돼지의 난포란을 체외에서 성숙, 수정시킨 체외수정란의 체외배양 체계를 확립하고 그 기작을 규명하기 위하여 체외배양액에 항산화제인 melatonin의 첨가 및 melatonin과 sodium nitroprusside(SNP)의 첨가배양이 체외수정란의 체외발육에 미치는 영향을 검토하고자 실시하였다. NCSU 23 배양액에 melatonin을 0, 1, 5 및 10nM을 첨가하여 체외배양을 실시한 결과, 배반포기까지 발육율은 17.8%, 26.1%, 20.0% 및 16.3%로서 melatonin 1nM 첨가구가 여타구에 비해 통계적으로 유의하게 높은 성적을 나타냈으며(P<0.05), 상실배기 이상 발육 성적에서도 melatonin 1 nM 첨가구가 39.1%로서 대조구 33.3%, 5 nM 첨가구의 33.3% 및 10 nM 첨가구의 27.9%보다 높은 발육율을 나타냈다(P<0.05). NCSU 23 배양액에 SNP를 0, 50 및 100 $\muM을 첨가하여 체외 배양한 결과, 상실배 이상 발육성적은 각각 41.9%, 25.6% 및 28.4%로서 SNP 첨가구가 대조구보다 유의적으로 낮은 성적을 나타내었다(P<0.05). NCSU 23 배양액에 대조구, SNP 50 $\muM, SNP 50 $\muM에 melatonin 1, 5 및 10nM을 혼합첨가하여 체외 발육율을 조사한 결과, 배반포기 발육율은 각각 2.5%, 1.2%, 9.9%, 5.1% 및 3.7%로서 SNP 50$\mu$M + Mel. 1nM 첨가구가 여타구 보다 높은 성적을 나타냈으며, 상실배기 이상 체외 발육율은 31.3%, 34.1%, 39.5%, 29.4% 및 39.5%로서 SNP 50 $\mu M + Mel. 1 nM 첨가구와 SNP 50 $\muM + Mel. 10 nM 첨가구가 여타구보다 높은 발육율을 나타냈다. 모든 처리구에서 배반포까지 발육된 체외수정란의 세포수는 커다란 차이가 인정되지 않았다.

• Archives of Pharmacal Research
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• 제16권1호
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• pp.1-5
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• 1993

The reaction of sulfhydryl groups in human serum ablumin with bacteriostatic and hypotensive notrosating agents such as sodium nitorprusside and sodium nitrite has been examined. The low reactivity of sodium nitroprusside to sulfhydral groups in albumin has been observed and the sterical inaccessilibility of the agent site which sulfhydryl group resides was implicated. The reaction of sodium nitrite with albumin was highly influenced by pH and little reactivity was observed at physiological pH. On the other hand, the reaction between albumin and S-nitrosoglutatione, an intermediate induced from the reaction of glutathione and nitrosating agents, resulted in the rapid decrease of free sulfhydryl groups in albumin. S-Nitrosylation of the sulfhydryl group by S-nitrosoglutathione and the subsequent production of mixed disulfide is the probable route of modification. In the physiological system, S-nitroso-glutathione may act as an active intermediate in expressing reacivity of nitrosating agents to sulfhydryl groups in albumin.

• Toxicological Research
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• 제16권2호
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• pp.133-139
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• 2000

This study was designed to evaluate the mechanism by which reactive nitrogen intermediates (RNI) induced the cytotoxicity of human doparminergic neuroblastoma SH-SY5Y cells. 3-Morpholino-sydnonimine (SIN-l), a donor of peroxynitrite (ONOO) and sodium nitroprusside (SNP), a donor of nitric oxide (NO) induced cell detachment and apoptotic death, as characterized by chromatin condensation, the ladder pattern fragmentation of genomic DNA and morphological nuclear changes. SIN-l also induced the activation of caspase 3-like protease in a time-dependent manner. Exogenous antioxidants, such as reduced glutathione (GSH), N-acetylcysteine (NAC), and selenium protected the cells from apoptotic death and reduced the activation of caspase 3-like protease by SIN-1. Furthermore, SIN-l directly reduced the intracellular levels of glutathione. Taken together, these data suggested that RNI including NO and peroxynitrite decrease the concentration of intracellular antioxidant such as GSH, which lead to the apoptotic death of human neuroblastoma SH-SY5Y cells.

• Journal of Ginseng Research
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• 제37권3호
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• pp.300-307
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• 2013

20S-dihydroprotopanaxatriol (2H-PPT) is a derivative of protopanaxatrol from ginseng. Unlike other components from Panax ginseng, the pharmacological activity of this compound has not been fully elucidated. In this study, we investigated the modulatory activity of 2H-PPT on the cellular responses of monocytes and macrophages to understand its immunoregulatory actions. 2H-PPT strongly upregulated the release of radicals in sodium nitroprusside-treated RAW264.7 cells and the surface levels of costimulatory molecule CD86. More importantly, this compound remarkably suppressed nitric oxide production, morphological changes, phagocytic uptake, cell-cell aggregation, and cell-matrix adhesion in RAW264.7 and U937 cells in the presence or absence of lipopolysaccharide, anti-CD43 antibody, fibronectin, and phorbal 12-myristate 13-acetate. Therefore, our results suggest that 2H-PPT can be applied as a novel functional immunoregulator of macrophages and monocytes.