ROLE OF PHOSPHOLIPASE C - δ1 IN OXYTOCIN RECEPTOR SIGNALING : EVIDENCE FOR AN EFFECTOR VIA Gh PROTEIN

백광진(Eu Suk Park),(Jong Hyun Won),(Pan Gil Suh),(Sung Ho Ryu),(Hee Sung Lee),(Hye Young Yun),(Nyoun Soo Kwon),(Kwang Jin Baek) 생화학분자생물학회 1997 생화학분자생물학회 춘계학술발표논문집 Vol.- No.-

Prednisone 투여에 의한 흰쥐 위점막 손상과 Liposomal Superoxide Dismutase의 보호효과

백광진,이희성,Baek, Kwang-Jin,Lee, Hee-Sung 생화학분자생물학회 1989 한국생화학회지 Vol.22 No.2

경구 투여방법으로 흰쥐에 prednisone (10 mg/100g body weight)을 투여하거나 liposomal SOD(3,000 units) 투여 후 prednisone(10 mg/100 g body weight)을 투여하여 1-4일 및 5일후에 위를 적출해서, 병리조직학적 소견과 위에서의 SOD 확인 및 SOD, catalase, peroxidase, xanthine oxidase 등의 활성도 변화를 관찰하여 다음과 같은 결과를 얻었다. 1. 대부분의 실험군에서, Cu, Zn-SOD, Mn-SOD, catalase의 활성도는 대조군보다 높았다. 2. Peroxidase의 활성도는, prednisone 투여 후 2일된 실험군만을 제외하고 모든 실험군이 대조군보다 유의하게 낮았다 (p<0.01). 3. Cu, Zn-SOD, Mn-SOD, catalase 및 peroxidase의 활성도는 prednisone만 투여한 실험군보다 liposomal SOD 투여 후 prednisone 투여한 실험군에서 높게 나타났다. 4. Xanthine oxidase의 활성도는 prednisone만 투여한 실험군이 liposomal SOD 투여 후 prednisone 투여한 실험군보다 항상 높았으며, prednisone만 투여한 실험군에서는 투여 후 2일만 제외하고 대조군보다 활성이 유의하게 높았다 (p<0.01). 5. KCN을 사용한 전기영동으로 Cu, Zn-SOD 및 Mn-SOD를 분리, 확인하였다. 6. Prednisone만 투여한 실험군에서는 위 병변 (정박 및 점막하 출혈, 궤사물)이 발생했으나, liposomal SOD 투여 후 prednisone 투여한 실험군에서는 위 병변이 발생하지 않았다. 이상과 같이 prednisone의 경구 투여로 xanthine oxidase의 활성이 증가되고 위 병변이 발생하였으나 liposomal SOD는 prednisone에 의한 위 병변을 효과적으로 억제했다. 그러므로 xanthine oxidase의 활성증가에 따른 superoxide radical과 그밖의 oxygen radical이 위점막 손상의 병인에 관계되리라 생각하며, SOD, catalase, peroxidase 등 oxygen radical을 제거하는데 관여하는 효소들은 위점막 방어기전에 중요한 역할을 할 것으로 생각한다. In the stomach of rats treated with prednisone (10 mg/100 g body weight) or liposomal superoxide dismutase (3,000 units) followed by prednisone (10 mg/100 g body weight), changes of the activities of superoxide dismutase (EC 1.5.1.1, SOD), catalase (EC 1.11.1.6), peroxidase (EC 1.11.1.7) and xanthine oxidase (1.1.3.22), as well as the macroscopical and microscopical changes within the gastric mucosa were investigated, and disc gel electrophoresis was used for confirmation of SOD in rat stomach. The results were summarized as follows: 1. In most of experimental groups, activities of Cu, Zn-SOD and catalase were higher than those in the controls. 2. In all experimental groups excepts 2nd day after the treatment of prednisone, the peroxidase activity showed lower than that in the controls (plt;0.01). 3. The activities of Cu, Zn-SOD, Mn-SOD, catalase and peroxidase in the liposomal SOD and prednisone treated groups appeared to be higher than those in the prednisone treated groups. 4. The activity of xanthine oxidase increased in the prednisone treated groups compared with the prednisone and liposomal SOD treated groups, and the activity showed higher in prednisone treated groups compared with the control value (plt;0.01). 5. Cu, Zn-SOD and Mn-SOD were confirmed by disc gel electrophoresis. 6. The gastric lesions (necrotic debris, mucosal and submucosal hemorrhage) occurred in the prednisone treated gorups, whereas, in the prednisone and liposomal SOD treated groups, the gastric mucosa were intact. From the above results, prednisone could enhance the activity of xanthine oxidase and results in gastric lesions, but the liposomal SOD may effectively protect the stomach from the development of gastric lesions. Accordingly, it could be suggested that the production of oxygen radicals (superoxide radical, hydroxyl radical, hydrogen peroxide and singlet oxygen) due to the increase of the activity of xanthine oxidase develop the gastric mucosal lesions. And SOD, catalase and peroxidase, scavengers of free radicals, may play an important role in the gastric mucosal defence mechanism.

Prednisone 투여에 의한 흰쥐 위점막 손상과 Liposomal Superoxide Dismutase 의 보호효과

백광진,이희성 ( Kwang Jin Baek,Hee Sung Lee ) 생화학분자생물학회 1989 BMB Reports Vol.22 No.2

In the stomach of rats treated with prednisone (10 ㎎/100 g body weight) or liposomal superoxide dismutase (3,000 units) followed by prednisone (10 ㎎/100 g body weight), changes of the activities of superoxide dismutase (EC 1.5.1.1, SOD), catalase (EC 1.11.1.6), peroxidase (EC 1.11.1.7) and xanthine oxidase (1.1.3.22), as well as the macroscopical and microscopical changes within the gastric mucosa were investigated, and disc gel electrophoresis was used for confirmation of SOD in rat stomach. The results were summarized as follows: 1. In most of experimental groups, activities of Cu, Zn-SOD and catalase were higher than those in the controls. 2. In all experimental groups excepts 2nd day after the treatment of prednisone, the peroxidase activity showed lower than that in the controls (p$lt;0.01). 3. The activities of Cu, Zn-SOD, Mn-SOD, catalase and peroxidase in the liposomal SOD and prednisone treated groups appeared to be higher than those in the prednisone treated groups. 4. The activity of xanthine oxidase increased in the prednisone treated groups compared with the prednisone and liposomal SOD treated groups, and the activity showed higher in prednisone treated groups compared with the control value (p$lt;0.01). 5. Cu, Zn-SOD and Mn-SOD were confirmed by disc gel electrophoresis. 6. The gastric lesions (necrotic debris, mucosal and submucosal hemorrhage) occurred in the prednisone treated gorups, whereas, in the prednisone and liposomal SOD treated groups, the gastric mucosa were intact. From the above results, prednisone could enhance the activity of xanthine oxidase and results in gastric lesions, but the liposomal SOD may effectively protect the stomach from the development of gastric lesions. Accordingly, it could be suggested that the production of oxygen radicals (superoxide radical, hydroxyl radical, hydrogen peroxide and singlet oxygen) due to the increase of the activity of xanthine oxidase develop the gastric mucosal lesions. And SOD, catalase and peroxidase, scavengers of free radicals, may play an important role in the gastric mucosal defence mechanism.

Azid 에 의한 Mouse 폐내 산소독성의 증강

백광진,권년수,이희성 ( Kwang Jin Baek,Nyoun Soo Kwon,Hee Sung Lee ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.2

An increased production of reactive oxygen species has been postulated to be a major factor in the pathogenesis of lung damage during hyperoxia. Azide is an inhibitor of superoxide dismutase (SOD) and catalase. These enzymes are important in defence mechanism against oxygen toxicity by removing superoxide radicals and hydrogen peroxide. Therefore, azide treatment could enhance oxygen toxicity. Mice were exposed to 100% O₂ (1 atm) with or without pretreatment with azide (8.1 ㎎/㎏ body weight). Biochemical indicators of oxygen toxicity in the lung of mouse were measured. Production of superoxide radicals and hydroxyl radicals increased in the lung of mouse treated with azide and 100% O₂. Superoxide radical generation was maximized at 24 h after the treatment, while the highest hydroxyl radical production occurred at 48 h. Xanthine oxidase activity was continuously decreased by the treatment with azide and 100% O₂. During the first 6 h of the treatment ∼90% of xanthine oxidase activity appeared as the type $quot;O$quot;. The Mn-SOD and glutathione peroxidase in azide and 100% O₂ treated group showed a significantly higher activity than those in the azide or 100% O₂ treated groups, while catalase activity was significantly lower in the azide/O₂-treated group. Survival time in azide and 100% O₂exposed group was significantly shorter than that in the azide or 100% O₂ treated groups. When SOD-containing liposomes (1,500 units/mouse) were introduced into the peritoneal cavity, the survival time was significantly extended. These results suggested that oxygen toxicity occurred in the lung of 100% O₂ exposed mouse by overproduction of reactive oxygen species and that the toxicity was exacerbated by azide-induced inhibition of enzymes in capable of removing reactive oxygen species. Liposomal SOD protected the mouse from the oxygen toxicity and extended the survival time.

흰쥐 폐에서 Prednisone 에 의한 산소 독성과 Liposomal Superoxide Dismutase 의 보호 효과

김수영,백광진,이희성 ( Soo Young Kim,Kwang Jin Baek,Hee Sung Lee ) 생화학분자생물학회 1990 BMB Reports Vol.23 No.3

The changes of superoxide dismutase (SOD), catalase, peroxidase and xanthine oxidase activities in the lung were observed after treating rats either with prednisone (10 ㎎/100 g body weight, group A) alone or with oral liposomal superoxide dismutase (3,000 units) followed by prednisone (10 ㎎/100 g body weight, group B). The results were summarized as follows: 1. In group A, Cu, Zn-SOD activity was higher than non-treated control values, but group B showed lower than that in the control. 2. The activity of Mn-SOD in group A increased progressively until the 3rd day after prednisone treatment and then decreased thereafter, but in group B, the activity was not changed except on the 4th day after the treatment. 3. In all experimental groups, the catalase activity was higher than that in the control, and gorup B showed higher activity than that in group A except on the 4th day after the treatment. 4. The activity of peroxidase in group A was higher than that in the control, but the activity in group B was similar to that in control. 5. In group A, xanthine oxidase activity showed higher than that in the control and group B except on the 1st day after the treatment, and the activity in group A increased with time. The above results suggest that prednisone administration could enhance the xanthine oxidase activity which lead to the production of oxygen radicals in the lung, and that the liposomal SOD may effectively protect the lung from the oxygen toxicity.

흰쥐 폐에서 Prednisone에 의한 산소 독성과 Liposomal Superoxide Dismutase의 보호 효과

김수영,백광진,이희성,Kim, Soo-Young,Baek, Kwang-Jin,Lee, Hee-Sung 생화학분자생물학회 1990 한국생화학회지 Vol.23 No.3

경구투여 방법으로 흰쥐에 prednisone(10mg/100g body weight)을 투여하거나(A 실험군) liposomal SOD(3,000 units) 투여 후 prednisone(10mg/l00g body weight)을 투여하여 (B 실험군) 1-4 및 5일 뒤에 폐를 적출해서 Cu, Zn-SOD, Mn-SOD, catalase, peroxidase, xanthine oxidase 등의 활성도 변화를 관찰하여 다음과 같은 결과를 얻었다. 1. Cu, Zn-SOD 활성도는, 대부분의 A 실험군이 대조군(생리 식염수만을 1ml/100g body weight 용량으로 경구투여)보다 증가했으나, B 실험군은 대조군보다 감소했다. 2. Mn-SOD 활성변화에서 A 실험군은 3일까지 활성이 점차 증가했으나 그 뒤에 활성이 감소했으며, B 실험군은 4일군만 제외하고 대조군과 비슷한 활성을 보였다. 3 모든 실험군에서 catalase의 활성도는 대조군보다 높았으며, 4일 군만 제외하고 B 실험군의 활성이 A 실험군보다 높았다. 4. Peroxidase의 활성변화에서, A 실험군 모두는 대조군보다 활성이 유의하게 증가한 반면, B 실험군 모두는 대조군과 비슷한 수준의 활성을 나타냈다. 5. Xanthine oxidase의 활성변화에서, A 실험군은 1일 군만 제외하고 대조군에 비해 활성이 유의하게 증가했으며, B 실험군과의 비교에서도 1일 군만 제외하고 활성이 항상 높았다. 또한 시간경과에 따라서도 활성이 증가하는 양상을 보였다. 이상과 같이 prednisone의 경구투여에 의해 xanthine oxidase의 활성이 증가되므로 superoxide radical 등 oxygen radical이 생성되어 폐에서 산소 독성 효과가 있으리라 생각하며, SOD, catalase, peroxidase 등 oxygen radical 제거에 관여하는 효소들은 활성이 증가하여 폐에서의 산소 독성 방어기전에 중요한 역할을 하는 것으로 생각한다. The changes of superoxide dismutase (SOD), catalase, peroxidase and xanthine oxidase activities in the lung were observed after treating rats either with prednisone (10 mg/100 g body weight, group A) alone or with oral liposomal superoxide dismutase (3,000 units) followed by prednisone (10 mg/100 g body weight, group B). The results were summarized as follows: 1. In group A, Cu, Zn-SOD activity was higher than non-treated control values, but group B showed lower than that in the control. 2. The activity of Mn-SOD in group A increased progressively until the 3rd day after prednisone treatment and then decreased thereafter, but in group B, the activity was not changed except on the 4th day after the treatment. 3. In all experimental groups, the catalase activity was higher than that in the control, and gorup B showed higher activity than that in group A except on the 4th day after the treatment. 4. The activity of peroxidase in group A was higher than that in the control, but the activity in group B was similar to that in control. 5. In group A, xanthine oxidase activity showed higher than that in the control and group B except on the 1st day after the treatment, and the activity in group A increased with time. The above results suggest that prednisone administration could enhance the xanthine oxidase activity which lead to the production of oxygen radicals in the lung, and that the liposomal SOD may effectively protect the lung from the oxygen toxicity.

사람 융모막 조직에서 Superoxide Dismutase 의 정제 및 성상

황윤영,김홍배,백광진,이희성 ( Yun Young Hwang,Hong Bae Kim,Kwang Jin Baek,Hee Sung Lee ) 생화학분자생물학회 1991 BMB Reports Vol.24 No.4

The cytosolic superoxide dismutase (SOD) in human chorion, which was obtained on normal delivery, was purified approximately 332-fold with 0.8% recovery. The enzyme was found to be a heterodimer composed of 39 kDa and 22 kDa subunits. The enzyme showed an activation at alkaline pH (10.0) and was inhibited by cyanide but not by azide, which suggests that the cytosolic SOD of human chorion is of a Cu, Zn-SOD type. The mitochondrial SOD of human chorion, which was purified 155-fold with 0.8% recovery, was found to be a homodimer composed of two 25 kDa subunits. In contrast to the cytosolic SOD, the mitochondrial SOD was markedly inhibited by the raise of pH in the reaction mixture from pH 7.8 to pH 10.0, and only slightly inhibited by cyanide and azide. The results suggest that the mitochondrial SOD of human chorion is of a Mn-SOD type as in other tissues.

사람 융모막 조직에서 Superoxide Dismutase의 정제 및 성상

황윤영,김홍배,백광진,이희성,Hwang, Yun-Young,Kim, Hong-Bae,Baek, Kwang-Jin,Lee, Hee-Sung 생화학분자생물학회 1991 한국생화학회지 Vol.24 No.4

정상 분만한 사람의 융모막 세포질내에 있는 superoxide dismutase(SOD)는 0.8%의 회수율로 332배 정제되었다. Sephadex G-100 gel filtration 및 HPLC에 의한 분자량은 61 kDa이었으며 SDS-PAGE에 의한 subunits의 분자량은 39 kDa, 22 kDa으로써 SOD는 두개의 다른 subunits으로 구성된 dimer 효소였다. 완충액의 pH가 10.0인 경우 효소의 활성이 3.5배 증가했으며, cyanide 농도에 비례해서 효소의 활성이 억제되었으며, azide는 효소 활성에 영향을 주지 못해서 Cu, Zn-SOD의 성상을 나타냈다. 융모막 mitochondria내에 있는 SOD는 0.8%의 회수율로 155배 정제되었다. Sephadex G-100 gel filtration 및 HPLC에 의한 효소의 분자량은 50kDa이었으며, SDS-PAGE에 의한 subunits의 분자량은 25 kDa으로써 두개의 동일한 subunits으로 구성된 dimer 효소였다. 완충액의 pH를 7.8에서 10.0으로 했을 때 효소의 활성이 억제되었으며, azide와 cyanide에 의해서 효소의 활성이 오직 조금만 억제되어 Mn-SOD의 성상을 나타냈다. The cytosolic superoxide dismutase (SOD) in human chorion, which was obtained on normal delivery, was purified approximately 332-fold with 0.8% recovery. The enzyme was found to be a heterodimer composed of 39 kDa and 22 kDa subunits. The enzyme showed an activation at alkaline pH (10.0) and was inhibited by cyanide but not by azide, which suggests that the cytosolic SOD of human chorion is of a Cu, Zn-SOD type. The mitochondrial SOD of human chorion, which was purified 155-fold with 0.8% recovery, was found to be a homodimer composed of two 25 kDa subunits. In contrast to the cytosolic SOD, the mitochondrial SOD was markedly inhibited by the raise of pH in the reaction mixture from pH 7.8 to pH 10.0, and only slightly inhibited by cyanide and azide. The results suggest that the mitochondrial SOD of human chorion is of a Mn-SOD type as in other tissues.

엑시머레이저 각막절제술 후 백내장 유발과 Nitric Oxide와의 관계

배노영(Noh Young Bae),류지환(Ji Han Ryon),백광진(Kwang Jin Baek),신경환(Kyung Hwan Shyn),김재찬(Jae Chan Kim) 대한안과학회 1998 대한안과학회지 Vol.39 No.11

진피세포의 조성이 인공피부의 기저막과 표피형성에 미치는 영향

이해란 ( Hailan Li ),정효순 ( Hyo-soon Jeong ),김잔디 ( Jandi Kim ),윤혜영 ( Hye-young Yun ),백광진 ( Kwang Jin Baek ),권년수 ( Nyoun Soo Kwon ),민영실 ( Young Sil Min ),박경찬 ( Kyoung-chan Park ),김동석 ( Dong-seok Kim ) 대한화장품학회 2012 대한화장품학회지 Vol.38 No.3

최근 유럽연합에서는 동물실험을 통하여 검증된 성분을 함유하고 있는 화장품에 대하여 판매를 금지할 것을 선언하였다. 그리하여 동물실험을 대체할 동물실험 대체모델의 개발이 필요해졌다. 인공피부는 화장품, 의약품 및 의료기기의 안전 테스트에 있어서 아주 중요한 시스템이다. 본 연구에서는 기저막과 표피를 가지고 있는 최적의 인공피부를 만들기 위한 시도를 하였다. 이러한 목적으로 중간엽줄기세표(MSCs, mesenchymal stem cells)와 지방전구세포를 진피세포인 섬유모세포와 혼합하여 진피대체물을 만들었다. 기저막과 표피의 형성은 면역조직화학 염색(immunohistochemical stains)을 통하여 확인하였다. 여러 가지 모델 중에서, 중간엽줄기세포를 혼합한 모델에서 표피의 두께가 제일 두꺼웠으며 또한 PCNA와 involucrin의 분포가 실제 사람피부와 비슷하였다. 결론적으로, 본 연구의 결과는 진피대체물에 중간엽줄기세포를 혼합한 인공피부가 동물실험 대체모델로 개발될 수 있다는 점을 제시한다. European Union prohibited the marketing of cosmetic products containing constituents that have been examined through animal experiments. Thus, non-animal test models are needed to replace animal experiments. The reconstructed skin models are important as a test system for cosmetic, pharmaceutical, and medical device safety testing. In the present study, we tried to develop an optimal skin equivalent model containing basement membrane and epidermis. For this purpose, we used mesenchymal stem cells (MSCs) and/or preadipocytes as well as fibroblasts as the dermal matrix cells. The formation of basement membrane and epidermis was verified by immunohistochemical stains. Among various models, the epidermis was thickest when MSCs were used in the dermal matrix. Furthermore, PCNA and involucrin distribution showed that dermal matrix with MSCs resembled human skin. Therefore, skin equivalents with MSCs could be developed as a non-animal test model to replace animal experiments.