백서와 기니픽의 대뇌피질에서 Opioid Kappa 수용체의 특성에 관한 연구

김기원(Kee-Won Kim),노혜원(Hye-Won Rho),김형일(Hyoung-Il Kim),은재순(Jae-Soon Eun),소수미(Soo-Mi Soh),조규박(Kyu-Park Cho) 대한약리학회 1994 대한약리학잡지 Vol.30 No.2

In this study, we tested the influences of several κ opioid ligands on the [³H]diprenorphine binding in rat and guinea pig cortex membrane preparations. Using paradigm to block μ and δ opioid receptors with DAMGO(1μM) and DPDPE(1μM), [³H]diprenorphine labeled κ sites. Competition analysis in both rat and guinea pig cortex has shown a single population of [³H]diprenorphine binding site with different Kd values, respectively. There is a significant difference in Ki values of (-) WIN44441 and (+)WIN44441 in both rat and guinea pig cortex. Bremazocine, (-)ethylketocyclazocine, (-)cyclazocine, nor-binaltorphimine effectively inhibited the [³H]diprenorphine binding with different Ki values in rat and guinea pig cortex. U-69,593, U-50,488H and dynorphine-A (1-8) did not inhibit the [³H]diprenorphine binding in rat but in guinea pig cortex. Nor-binaltorphimine was a ligand discriminate the κ₁, and κ₂ receptor most effectively. We, also, examined the influence of Na ion and GTPγS, a nonhydrolyzable guanine nucleotide analog, on the inhibition of [³H]diprenorphine binding by diprenorphine, (-)ethyl-ketocyclazocine, U-69,593 and bremazocine. By the replacement of NaCl with N-methy-D-glucamine or addition of GTPγS, Ki values of diprenorpnine were not changed and that of ethylketocyclazocine were changed significantly in both rat and guinea pig cortex. The Ki value of bremazocine was decreased by removal of Na ion, and increased by GTPγS, however, was not changed by any one of either. These results suggest that there are 2 kinds of subtypes of κ opioid receptor, κ₁, and κ₂, showing different Ki values for various κ opioid ligands, also, bremazocine possess the antagonistic property at κ₂ site which is dominant subtype of K receptor in rat cortex.

원저 : 마우스 소장조직 (小腸組織)의 Ornithine Aminotransferase 에 관한 연구

노혜원(Hye Won Rho),최경수(Kyung Soo Choi),임방은(Bang Eun Lim),김종석(Jong Suk Kim),박병현(Byung Hyun Park),김형로(Hyung Rho Kim) 전북대학교 의과학연구소 1991 全北醫大論文集 Vol.15 No.4

백일해 독소에 의한 중성 백혈구 chemotaxis 의 억제에 관한 연구

김정수,김우현,노혜원,김종석,박진우,김형로 ( Jung Soo Kim,Uh Hyun Kim,Hye Won Rho,Jong Suk Kim,Jin Woo Park,Hyung Rho Kim ) 생화학분자생물학회 1989 BMB Reports Vol.22 No.2

It has been suggested that pertussis toxin (PT) inhibits the chemotaxis of neutrophil by modifying signal transduction system coupled to phospholipase C. The PT substrate, probably the inhibitory guanine nucleotide-binding regulatory component of adenylate cyclase (Gi) or an analogous protein, is hence proposed to mediate N-formyl-methionyl-leucyl-phenylalanine (fMLP) receptor-linked activation of the phospholipase C. The aim of this investigation was to determine whether the role played by PT in this system, is dependent upon the ability of the A-protomer of the toxin to ADP-ribosylate Gi or secondary to the binding of the B-oligomer of the toxin to the surface of target cells. PT and methylated PT (mPT) were employed in parallel to assess the involvement of two distinct biologic activities of the toxin in the inhibition of chemotaxis and its related events. Our preparation of mPT retained its ability to ADP-ribosylate Gi, but the pretreatment of mPT to neutrophils did show the inhibition of the chemotaxis as well as the other events related to the phospholipase C, indicating that the involvement of PT is not solely dependent on its capacity to ADP-ribosylate Gi, but rather related to the role of B-oligomer of the toxin.

The Inhibition of Neutrophil Chemotaxis by Pertussis Toxin

김정수,김우현,노혜원,김종석,박진우,김형로,Kim, Jung-Soo,Kim, Uh-Hyun,Rho, Hye-Won,Kim, Jong-Suk,Park, Jin-Woo,Kim, Hyung-Rho 생화학분자생물학회 1989 한국생화학회지 Vol.22 No.2

백일해 독소(PT)는 phospholipase C와 연결된 신호전달 체계를 변형시켜 중성 백혈구의 chemotaxis를 억제한다고 알려져 왔다. fMLP 수용체에 연결되어 phospholipase C가 활성화 되는데는 adenylate cyclase 체계를 억제하는 guanine nucleotide의 결합 조절 단백질(Gi 단백질) 또는 상동성 단백질이 신호전달을 중계할 것이라고 생각되고 있다. 본 연구에서는 이 체계에 대한 PT의 역할이 A-protomer의 ADP-ribosyltransferase 작용에 의한 Gi 단백질의 변형 때문인지 또는 B-oligomer의 세포 표면 결합에 의한 이차적인 효과인지를 구명하였다. PT와 메틸화 PT(mPT)는 chemotaxis 억제 및 관련된 현상들에 서로 다른 생물학적 활성을 가진 독소의 두 부분중 어느 부분이 관여하는지를 알기 위하여 이용하였다. 조제한 mPT는 ADP-ribosyltransferase 활성을 유지하고 있음을 확인하였고 이 mPT는 chemotaxis 억제능력이 없음을 관찰함과 동시에 그에 수반되는 생화학적 사건들에서도 일관성 있는 결과를 얻었기에 PT에 의한 chemotaxis의 억제는 A-protomer보다는 B-oligomer의 역할과 관계되리라 사료된다. It has been suggested that pertussis toxin (PT) inhibits the chemotaxis of neutrophil by modifying signal transduction system coupled to phospholipase C. The PT substrate, probably the inhibitory guanine nucleotide-binding regulatory component of adenylate cyclase (Gi) or an analogous protein, is hence proposed to mediate N-formyl-methionyl-leucyl-phenylalanine (fMLP) receptor-linked activation of the phospholipase C. The aim of this investigation was to determine whether the role played by PT in this system, is dependent upon the ability of the A-protomer of the toxin to ADP-ribosylate Gi or secondary to the binding of the B-oligomer of the toxin to the surface of target cells. PT and methylated PT (mPT) were employed in parallel to assess the involvement of two distinct biologic activities of the toxin in the inhibition of chemotaxis and its related events. Our preparation of mPT retained its ability to ADP-ribosylate Gi, but the pretreatment of mPT to neutrophils did show the inhibition of the chemotaxis as well as the other events related to the phospholipase C, indicating that the involvement of PT is not solely dependent on its capacity to ADP-ribosylate Gi, but rather related to the role of B-oligomer of the toxin.

사인의 항당뇨 작용에 관한 연구

이지현(Ji Hyun Lee),조정임(Zheng Lim Zhao),조남표(Nam Pyo Cho),박병현(Byung Hyun Park),권강범(Kang Beom Kwon),노혜원(Hye Won Rho) 한의병리학회 2007 동의생리병리학회지 Vol.21 No.2

Virbrio Vulnificus가 분비하는 Cytolysin의 성상

노혜원,김종석,양병철,최원용,김형로 의과학연구소 1989 全北醫大論文集 Vol.13 No.1

The cytolysin produced by Vibrio vulnificus was isolated and its properties were studied. 1. The cytolytic activity in the supernatant of culture media reached to the maximum about 4-5 hours of culture and decreased rapidly thereafter. Meanwhile, the protease activity increased with the time until 8 hours of culture and maintained during culture period of 12 hours. 2. The cytolysin was lethal for mouse and its LD^50 for mouse ws estimated as 3.7 hemolytic units. 3. The cytolysin was very active against sheep and mouse erythrocytes, while it was less active against rabbit and human erythrocytes. 4. The cytolytic activity was maximum in the presence of 0.5mg/ml BSA in assay mixture. (Key words : Vibrio vulnificus, cytolysin, protease)

마우스 小腸組織의 Ornithine Aminotransferase에 關한 硏究

노혜원,최경수,임방은,김종석,박병현,김형로 의과학연구소 1991 全北醫大論文集 Vol.15 No.4

Ornithine aminotransferase catalyzes the transfer of δ-amino group L-ormithine to α-ketoglutarate to yield glutamic- γ - sernialdehyde and glutamate. In present study, ornithine aminotransferase from mouse small intestine was puriried and its biochemical properties were studied. 1.Ornithine aminotransferase activity ws higher in mouse small intestine than in liver and kidnery. The enzyme ws found to be mostly located in cytosolic fraction of small intestine, but present in mitochondrial fraction of liver and kidney. 2.The cytosolic enzyme was very unstable, but the stabiling was enhanced by the presence of EDTA and glycerol. 3.The cytosolic ornithine aminotransferase was purified to homogeniety from mouse small intestine by heat treatment, calcium-phosphate gel adsorption, DEAE= cellulose chromatography, Sephadex G-100 gel filtration and Protein Analysis I-125 gel filtration. The specific a activity of 240-fold purified enzyme was 806 unit/mg protein. 4.The purified enzyme is composed of a single polypeptide chain with molecular weight of 46,000-47,000 daltons and its optimum pH was 8.5. The isoelectric point of the enzyme was found to be pH 5.3, suggesting that the content of acidic amoino acids is relatively high in the enzyme protein.

알록산 투여로 유발된 실험적 당뇨병에 대한 사인추출물의 치료 효과

노혜원,이지나,구본선,조정임,박진우,김형로 대한당뇨병학회 2002 Diabetes and Metabolism Journal Vol.26 No.2

연구배경:사인추출물이 알록산에 의한 당뇨 유발을 억제하는 효과가 있음을 이미 확인하였던 바 사인 추출물이 췌장 베타세포가 손상되어 유발된 당뇨병의 치료에 사용할 수 있는지의 가능성을 알아보았다. 방법:실험적 당뇨는 알록산(60㎎/㎏)을 생쥐 꼬리 정맥을 통해 주사하여 유발시켰다. 당뇨에 미치는 사인추출물의 영향을 관찰하기 위하여 사인추출물의 영향을 관찰하기 위하여 사인추출물(2.5㎎/생쥐)을 하루에 두 번 생쥐 복강 내로 투여하였다. 당뇨에 미치는 사인추출물의 영향을 관찰하기 위하여 생쥐에 사인추출물을 처리한 다음 혈당 및 혈청 인슐린 농도 변화 및 조직학적인 소견으로 확인하였다. 결과:생쥐에 알록산을 투여하면 췌장 베타세포가 파괴되어 고혈당 및 hypoinsulinemia가 유발되었다. 사인추출물을 알록산 투여 전 2일 동안 전처리하면 알록산에 의한 고혈당이 완전히 억제되었다. 뿐만 아니라 알록산에 의해 유발된 췌장 베타세포의 손상은 사인추출물의 투여로 현저히 개선되어 혈당 및 혈청 인슐린 농도가 거의 정상 수준으로 회복되었다. 알록산을 투여한 생쥐의 췌장 소도는 정상군에 비해 수가 현저히 감소되었을 뿐 아니라 남아있는 소도의 크기도 현저히 줄어들어 있었으나, 사인추출물을 처리하면 모두 회복되어 정상군과 유사한 양상을 보였다. 결론:사인추출물은 알록산에 의해 유발되는 당뇨병을 억제하는 방어 작용 뿐만 아니라, 알록산 투여로 이미 손상된 췌장 베타세포를 개선시킬 수 있는 치료 효과작용도 가지고 있음을 확인하였다. 이를 토대로 사인의 당뇨 개선 기전을 규명하면 당뇨병 치료제로서 이용할 수 있으리라 사료된다. Background : During the screening of natural products for potential antidiabetogenic components, a strong protective effect of Amomum xanthoides extract on alloxan-induced β-cell damage and in a mice diabetic model. In this study, the therapeutic effect of Amomum xanthoides extract was investigated after induction of diabetes by alloxan. Methods : Experimental diabetes was induced by the injection of alloxan(60mg/kg) to the mouse via the tail vein. To examine the effect the of Amomum xanthoides extract on diabetes, Amomum xanthoides extract (2.5mg/mouse) was admini-strated intraperitoneally. The effect of the Amomum xanthoides extract on alloxan-induced diabetes was observed by measuring the blood glucose and serum insulin level, and a histological examination. Results : Alloxan caused hyperglycemia and hypoinsulinemia by a selectively destroying pancreatic β-cell. Pretreating the with an Amoumum xanthoides extract completely protected them from the hyperglycemia induced by alloxan. In addition, the Amomum xanthoides extract administe 3 days after the of alloxan injection significantly abolished the hyperglycemia and hypoinsulinemia induced by alloxan. The alloxan-treated mice showed a marked change of in the pancreatic islets: the number of islets was reduced and the size of the remaining islets also decreased. However these effects of alloxan were significantly recovered by a later administration of the Amomum xanthoides extract. Conclusion : The amomum xanthoides extract contains potentially effective components, which both protect and treat alloxan-induced diabetes. The identification and action mechanism of the effective components of the Amomum xanthoides extract requires further investigation and it is suggested that the Amomum xanthoides extract be used as a therapeutic drug for diabetes(J Kor Diabetes 26:126~133, 2002).

사람 소장 Ornithine Aminotransferase 의 성상

지은정,노혜원,양병철,조옥현,김형로 의과학연구소 1989 全北醫大論文集 Vol.13 No.1

Some properties of ornithine aminotransferase, which transfers the amino group of ornithine to α-ketoglutarate in human intestine were characterized. The results of this study were as follows. 1.Ornithine aminotransferase showing optimal pH 8.5 was mainly localized in cytosol and its Km for L-ornithine and α-ketoglutarate was 2.0 mM and 2.7 mM respectively. The enzyme exhibited the absolute requirement of pyridoxal phosphate for its activity. 2.By storage at 4℃for 5 day, 75% of the enzyme activity was lost and on heat treatment at 60℃ for 5 min, 80% of that was inactivated but the stability was markedly increased to almost 100% by the addition of 0.05mM pyridoxal phosphate to the enzyme solution. 3. From the fact that the enzyme activity is completely inhibited by the presence of p-chloromercuribenzoate or HgCl_2, it can be strongly suggested that sulfhydryl group is involved in the catalytic site of ornithine aminotransferase.

Lanthanum Chloride의 抗糖尿 效果

金炯魯,盧惠援,朴鎭宇 전북대학교 의과학연구소 1984 全北醫大論文集 Vol.8 No.2

Since Ca^2+ plays many important roles in cellular metabolism, an attempt was made to investigate the effect of lanthanum, a Ca^2+-antagonist, on alloxan- or streptozotocin-induced diabetes mellitus. Alloxan(80mg/kg bcdy weight) induced permanent hyperglycemia in rats reaching the blood glucose level of 585mg% at 48 hours and about two-tbird of rate was died for 5-day period after alloxan treatment. Lanthanum chloride(100mg/kg body weight) administration 1 hour before alloxan comepletely prevented alloxan-induced hyperglycemia, while it had no effect on streptozotocin-induced hyperglycemia, suggesting that lanthanumacts on a certain step between alloxan uptake into B-cell and the formation of free radicals plausible causative factor of alloxan-diabetogenesis. Lanthanum treatment at 24 hours after alloxan administration partially prevented alloxan-induced hyperglycemia, indicating that lanthanum stimulates the recovering process of B-cell damaged by alloxan.