거식세포 Nitric Oxide 생성의 생화학

권년수 중앙대학교 의과대학 의과학연구소 1991 中央醫大誌 Vol.16 No.1

Synthesis of nitric oxide is a recently discovered property of macrophages, endothelial cells, neutrophils, hepatocytes, and cells in adrenal gland and cerebellum. In all cell types, L-arginine is utilized to produce nitric oxide, leaving L-citrulline. Nitric oxide mediates some of cytotoxic actions of macrophages. Endothelium-derived nitric oxide is a major relaxant of vascular smooth muscle. In macrophages, nitric oxide synthase activity is induced by immunologic stimuli. The activity is cytosolic, exhibits a strict requirement for NADPH. Nitric oxide synthase requires tetrahydrobioptein as a cofactor, and reductases for reduced biopterins(e.g., dihydropteridine reductase) are necessary for continuous recycling of the cofactor. FAD and a reduced thiol such as glutathione are necessary for maximal nitric oxide generation by nitric oxide synthase. Stable isotope studies have shown that nitric oxide derives form one of the two equivalent guanidino nitrogens of L-arginine, and the dioxygen is the source of the oxygens incorporated into L-citrulline and nitric oxide. Nitric oxide syntheses is initiated by generation N^ω-hydroxy-L-arginine as an intermediate through an NADPHdependent hydroxylation of L-arginine. Nitric oxide synthase exclusively utilized the hydroxylated guanidino nitrogen for nitric oxide synthesis. Further reaction of N^ω-hydroxy-L-arginine to form nitric oxide and L-citrulline requires oxidations of NADPH and tetrahydrobiopterin.

Electroconvulsive Shock이 마우스 뇌조직 Superoxide Dismutase의 활성에 미치는 영향

권년수,이희성,Kwon, Nyoun-Soo,Lee, Hi-Sung 생화학분자생물학회 1984 한국생화학회지 Vol.17 No.3

In the present study, prolonged effects of the serial electroconvulsive shocks on the activity of superoxide dismutase (superoxide: superoxide oxidoreductase, EC 1.15.1.1): both cyanide-sensitive cytosolic enzyme and cyanide-resistant mitochondrial enzyme, were observed in the mouse brain. The rate constants for synthesis and degradation of these enzymes were determined. In addition, the changes in the superoxide radical production, the amount of hydrogen peroxide and the activity of superoxide dismutase were observed in the mouse brain which received single electroconvulsive shock. The results were summarized as follows: 1. Homogenates prepared from the brains of mice treated with single electroconvulsive shock showed considerably higher superoxide radical production when compared with those of the control mice. 2. By single electroconvulsive shock the generation of hydrogen peroxide increased at 1 minute after the electric stimulus; while the activity of superoxide dismutase increased at the time when tonic convulsion occurred. 3. By the serial electroconvulsive shocks the activity of cytosolic superoxide dismutase increased to approximately 1.6 times its control value ($Kd=3.60{\times}10^{-2}/hr$; Ks=6.01 units/hr/g tissue; $t_{1/2}=19\;hrs$); and the mitochondrial enzyme increased to approximately 2 times its control value ($Kd=1.53{\times}10^{-2}/hr$; Ks=1.44 units/hr/g tissue; $t_{1/2}=45\;hrs$). 4. The rate constants at recovery period were: $Kd=1.91{\times}10^{-2}/hr$; Ks=2.01 units/hr/g tissue; $t_{1/2}=36\;hrs$ for the cytosolic superoxide dismutase and $Kd=3.08{\times}10^{-2}/hr$; Ks=1.42 units/hr/g tissue; $t_{1/2}=23\;hrs$ for the mit ochondrial enzyme. ICR순계 마우스에 electroconvulsive shock(ECS)을 가해 뇌조직에서 superoxide radical의 생성, $H_2O_2$의 양, superoxid edismutase(superoxide: superoxide oxidoreductase, EC 1.15.1.1)의 활성에 미치는 영향 등을 관찰하고, 주기적으로 반복하여 ECS를 가함으로써 superoxide dismutase(SOD)를 유발시켜 합성 및 분해속도상수를 구한 결과 다음과 같은 결론을 얻었다. 1. ECS를 받은 마우스의 뇌조직 균질액에서 superoxide radical의 생성이 대조군에 비해 유의한 차이가 있게 증가되었다(p<0.01). 2. ECS에 의해 마우스 뇌조직의 SOD 활성도는 긴장성 경련기에, $H_2O_2$의 양은 1분후에 가장 높은 증가를 나타내었으며 SOD 활성도와 $H_2O_2$양의 변화 양상은 비슷하였다. 3. 일련의 ECS로 세포질 SOD는 대조군의 약 1.6배로 전기자극 후 11일에 가장 높은 활성을 나타냈다($Kd=3.60{\times}10^{-2}/hr$; Ks=6.01 units/hr/g tissue; $t_{1/2}=19\;hrs$). Mitochondria의 SOD는 전기자극후 11일에 대조군의 약 2배로 가장 높은 활성을 나타내었다($Kd=1.53{\times}10^{-2}/hr$; Ks=1.44 units/hr/g tissue; $t_{1/2}=45\;hrs$). 4. 회복기에 세포질 효소의 속도상수는 Kd가 $1.91{\times}10^{-2}/hr$, Ks가 2.01 units/hr/g tissue, 반감기는 36시간이었으며, mitochondria 효소의 Kd는 $3.08{\times}10^{-2}/hr$, Ks는 1.42 units/hr/g tissue, 반감기는 23시간이었다.

Electroconvulsive Shock 이 마우스 뇌조직 Superoxide Dismutase 의 활성에 미치는 영향

권년수,이희성 ( Nyoun Soo Kwon,Hi Sung Lee ) 생화학분자생물학회 1984 BMB Reports Vol.17 No.3

In the present study, prolonged effects of the serial electroconvulsive shocks on the activity of superoxide dismutase (superoxide: superoxide oxidoreductase, EC 1.15.1.1) : both cyanide-sensitive cytosolic enzyme and cyanide-resistant mitochondria) enzyme, were observed in the mouse brain. The rate constants for synthesis and degradation of these enzymes were determined. In addition, the changes in the superoxide radical production, the amount of hydrogen peroxide and the activity of superoxide dismutase were observed in the mouse brain which received single electroconvulsive shock. The results were summarized as follows: 1. Homogenates prepared from the brains of mice treated with single electroconvulsive shock showed considerably higher superoxide radical production when compared with those of the control mice. 2. By single electroconvulsive shock the generation of hydrogen peroxide increased at 1 minute after the electric stimulus; while the activity of superoxide dismutase increased at the time when tonic convulsion occurred. 3. By the serial electroconvulsive shocks the activity of cytosolic superoxide dismutase increased to approximately 1.6 times its control value (Kd=3.60×10^(-2)/hr; Ks=6.01 units/hr/g tissue ; t_½=19 hrs) ; and the mitochondriaI enzyme increased to approximately 2 times its control value (Kd=1.53×10^(-2)/hr; Ks=I.44 units/hr/g tissue; t_½=45 hrs). 4. The rate constants at recovery period were: Kd=1.91×10^(-2)/hr;Ks=2.01 units/hr/g tissue; t_½=36 hrs for the cytosolic superoxide dismutase and Kd =3.08×10^(-2)/hr; Ks=1.42 units/hr/g tissue; t_½=23 hrs for the mit ochondrial enzyme.

사람 태반의 Superoxide Dismutase의 정제 및 성상

문진수,권년수,이근배,이희성 중앙대학교 의과대학 의과학연구소 1982 中央醫大誌 Vol.7 No.2

The distribution and some properties of superoxide dismutase of human term placenta have been studied. Human placenta was fractionated by differential centrifugation into mitochondrial and cytosolic fractions. The activity of superoxide dismutase was measures by the methodof McCord and Fridovich. Cytosolic superoxide dismutase was purified by ammonium sulfate precipitation, treatment with a chloroform-ethanol mixture, and DEAE-cellulose column chromatography. The results are summarized as follows; 1. Human term placenta contains two types of superoxide dismutase, one of which is localized in the mitochondria while the other is found in the cytosol. The mitochondrial superoxide dismutase was inactivated by treatment with a moxture of chloroform and ethanol whereas the cytosolic superoxide dismutase was not. 2. The activity of cytosolic and mitochondrial superoxide dismutase was found to be 7.44 units and 1.27 units per g of wet tissue, repectively. 3. The molecular weight of cytosolic superoxide dismutase was estimated to be about 33,000 by gel filtration. 4. The ultraviolet absorption spectrum of cytosolic enzyme indicates the lack of tryptophan. The spectrum of the enzyme in the uitraviolet region was similar to that of phenylalanine. 5. Purified cytosolic superoxide dismutase contains 2Cu^2+ and 2Zn^2+ per molecule. This enzyme was found to be similar to the other cupro-zinc superoxide dismutase which have been isolated from diverse eukaryotes. 6. Cyanide at 0.5 mM and 5.0 mM inhibits the activity of cytosolic superoxide dismutase 31% and 98%, respectively, but mitochondrial superoxide dismutase was not inhibited by this compound.

정상 및 Phenylhydrazine을 처리한 토끼적혈구의 Isocitrate Dehydrogenase Isozymes 활성도에 관한 연구

정동철,권년수,이동욱,이희성 중앙대학교 의과대학 의과학연구소 1982 中央醫大誌 Vol.7 No.3

In order to establish a general pattern of synthesis and degradation of cytosolic and mitochondrial isocitrate dehydrogemase isozymes in the red blood cells of normal and phenlydeazine treated rabbits were conducted in this study. The activity of isocitrate dehydrogemase was measured by the method of King. The results and summarized as follows: 1. The reticulocyte count of normal rabbit blood was approximately 2.5%, and the administration of phenlydeazine resulted in rapid and profound reticlocytosis, i.e., about 63.5% on dat essentially 100% on day 7. 2. Total activity of isocitrate dehydrogenase in normal blood cells was 0.95 unit/ml RBC and the relative activities of cytosolic and mitochondrial isocitrate dehydeogenase isozymes were 91.6% and 8,4%, respectively. 3. In phenlydeazine treated rabbits, total isocitrate dehydrogenase activity was maximal on day 8, about 2.57 unit/ml RBC, which is about 2.7 fold higher than normal value. The relative activities of cytosolic and mitochondrial isocitrate dehydrogenase were 56.8% and 43.2%, respectively. 4. The increase in total isocitrate dehydrogenase activity during the induction of reticulocytisis appeared to be primarily due to 14.3 fold increase in the activity of the mitochondrial isocitrate dehydrogenase isozyme(Ks=0.0152; Kd=0.0133; t1/2=52.11 hrs). 5. The activity of the cytosolic isocitrate dehydrogenase isozyme in phenylhydrazine treated rabbit red blood cells increase to about 1.7 times its normal value(Ks=0.0102;Kd=0.0070; t1/2=99.00 hrs). 6. The rate constants following recovery from phenylhydrazine treatment were: Ks=0.0008; Kd=0.0104;t1/2=66.63 hrs for the mitochondrial isozyme and Ks=0.0208; Kd=0.0239; t1/2=29.00 hrs for the cytosolic isozyme.

허혈-재관류에 의한 흰쥐 간의 손상에 있어서 반응성 산소종의 역할

지경천,권년수,이희성,김상준,장선택 중앙대학교 의과대학 의과학연구소 1992 中央醫大誌 Vol.17 No.4

Increased generation of reactive oxygen species is thought to be a major cause of tissue in ischemia-reperfusion injury. Left and median lobes of rat livers were subjected to 20 minutes of ischemia followed by reperfusion for up to 120 minutes. Superoxide radical generation was enhanced to 338 μmol/minute/g tissue after 60 minutes of reperfusion,and 367 μmol/minute/g tissue after 120 minutes. Superoxide generation in the liver of the control animal was 260 μmol/minute/g tissue. The activity of xanthine oxidase, a superoxide radical-generating enzyme, was also elevated in the liver treated with ischemia-referfusion. These results indicate that xanthine oxidase-catalyzed reaction is an important source for the superoxide generation in rat liver treated with ischmia-referfusion. The activities of superoxide dismutase and catalase, enzyme responsible in removing superoxide radical and hydrogen peroxide, were measured. Cu, Zn-superoxide dismutase activity was significantly higher in reperfused liver after ischemia than in control. However, the activity of Mn-containing superoxide dismutase was significantly low in reperfusion. The extent of tissue damage by ischemia-reperfusion was evaluated by measuring lipid peroxidation. Thiobarbituric acid reactive substance was significantly elevated in the liver treated with 60 minutes of ischemia followed by 60 minute of reperfusion. These results suggested that increased reactive oxygen species and diminished activities of enzyme removing toxic oxygen species rendered the damages on the liver which was sujected to ischemia followed by reperfusion.

The role of nitric oxide in ocular surface diseases

Gun Sic Park,Nyoun Soo Kwon,Young Myeong Kim,Jae Chan Kim 대한안과학회 2001 Korean Journal of Ophthalmology Vol.15 No.2

The role of nitric oxide (NO) in ocular surface diseases remains unknown. We investigated the conditions leading to increase NO generation in tears and the main sources of ocular surface tissue. We evaluated the possibility of a dual action (cell survival or cell death) depending on the amount of NO. The concentration of nitrite plus nitrate, the stable end-product of NO, was measured in the tears of various ocular surface diseases. We also examined the main source of nitric oxide synthase (NOS) using immunohistochemical staining & Western blot analysis. When cultured human corneal fibroblasts were treated with NO producing donor with or without serum, the viability of cells was studied. We found that sources of NO in ocular surface tissue primarily included corneal epithelium, fibroblasts, endothelium and inflammatory cells. Three forms of NOS (eNOS, bNOS, & iNOS) were expressed in experimentally induced inflammation. Cell death by NO revealed TUNEL positive staining, however in the EM finding, this NO specific cell death was an atypical necrosis showing perinuclear large vacuolization and mitochondrial swelling. In the fibroblasts culture system, the NO donor (SNAP, S-nitroso-N-acetyl-D, L-penicillamine) prevented the death of corneal fibroblasts caused by serum deprivation in a dose dependent manner up to 500(m SNAP, although a higher dose decreased cell viability. This study suggested that NO might act as a double-edged sword in ocular surface disease depending on the degree of inflammatory condition related with NO concentration.

Leucine-rich glioma inactivated 3: Integrative analyses support its role in the cytokine network

Kim, Hyun A,Kwon, Nyoun Soo,Baek, Kwang Jin,Kim, Dong-Seok,Yun, Hye-Young Spandidos Publications 2017 International journal of molecular medicine Vol.40 No.1

<P>Leucine-rich glioma inactivated (LGI)3 is a secreted protein member of LGI family. We previously reported that LGI3 was upregulated in adipose tissues from obese mice and suppressed adipogenesis through its receptor, a disintegrin and metalloproteinase domain-containing protein 23 (ADAM23). We demonstrated that LGI3 regulated tumor necrosis factor- and adiponectin, and proposed that LGI3 may be a pro-inflammatory adipokine involved in adipose tissue inflammation. In this study, we analyzed adipokine and cytokine profiles in LGI3 knockout mice and demonstrated that multiple factors were increased or decreased in the adipose tissues and plasma of the LGI3 knockout mice. Phosphoprotein array analysis revealed increases in the phosphorylation levels of Akt, AMP-activated protein kinase (AMPK), Bad, extracellular signal-regulated kinase (Erk)1/2, glycogen synthase kinase 3 (GSK3), phosphatase and tensin homolog (PTEN) and eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) in the LGI3-treated 3T3-L1 pre-adipocytes. Treatment with LGI3 increased the expression of various inflammatory genes in pre-adipocytes, adipocytes and macrophages. Integrative functional enrichment analysis for all LGI3-regulated gene products suggested their involvement in a number of biological processes, including cancer, inflammatory response, response to wounding, as well as cell proliferation and differentiation. Protein interaction network analysis of LGI3-regulated gene products revealed that 94% of the gene products formed a cluster of interaction networks. Taken together, these results support the critical involvement of LGI3 in the cytokine network by interplaying with multiple adipokines, cytokines and signaling proteins.</P>

Gα^(h)-단백질에 의해 활성화되는 Phospholipase C 이소효소의 확인

조강호,박권식,채규영,한기정,김형석,김형일,윤혜영,권년수,이희성,백광진 중앙대학교 의과대학 의과학연구소 2000 中央醫大誌 Vol.25 No.4

Effects of Nitric Oxide on the Growth of Salmonella paratyphi B

Cheol Kyu Lee,Nyoun Soo Kwon,Chul Yong Song,Hee Sung Lee 생화학분자생물학회 1993 생화학분자생물학회 춘계학술발표논문집 Vol.- No.-