Cloning of the Structural Sequences of Human Growth Hormone Gene Family into Plasmids and Its Utilization in Transient Expression Systems

남궁욱,김용주,이경광,한문희,김지영,NamGung, Uk,Kim, Yong-Joo,Lee, Kyung-Kwang,Han, Moon-H.,Kim, Ji-Young 생화학분자생물학회 1988 한국생화학회지 Vol.21 No.3

사람 성장홀몬(hGH) 유전자와 코리온 소마토마모트로핀-A형과 B형 유전자의 구조 부위를 유전자농축방법에 의해 제조된 pBR322 사람 유전자 라이브러리로부터 분리하였다. 클로닝된 유전자들은 제한효소 지도 및 써던 하이브리다이제이션 분석에 의해 확인되었다. 반복되는 염기서열 (Repeated sequences)의 DNA 부위가 hGH 유전자의 3' 아랫쪽에 포함되어 있으나 hCS-A와 B형 유전자에는 있지 않았다. hGH 구조 유전자를 생쥐 메탈로티오닌-I(MT-I) 프로모터에 연결하여 MT-I 프로모터/hGH 융합 유전자가 포함된 재조합 플라스미드, pMThGH를 제조하였다. 이 플라스미드를 생쥐 L 세포에 넣어주었을 때 사람 성장홀몬은 MT-I 프로모터 조절하에 발현되었다. 사람 성장홀몬은 배양액으로 분비되었으며 약 10일간 생성이 지속되었다. 카드뮴으로 MT-I 프로모터를 유도하였을때 사람 성장홀몬의 농도는 몇배 증가하였다. 본 실험 결과는 사람 성장홀몬 구조 유전자가 클로닝된 플라스미드 벡터는 유전자 발현 조절 연구를 하는데 일시적인 발현 시스템에서 이용될 수 있음을 보여주었다. The structural sequences of human growth hormone(hGH), human chorionic somatomammotropin(hCS)-A, hCS-B genes were isolated from a partial human genomic DNA library of pBR322. The cloned genes were identified by the restriction and Southern blot hybridization analysis. A block of highly repeated sequences was contained In the 3'-flanking sequences of hGH gene but not in the hCS-A and hCS-B genes. The structural sequences of hGH gene was fused with mouse metallothionein-I(MT-I)promoter and the expression plasmid DNA was. introduced into mouse L cells by transfection. hGH was transiently expressed in the transfected mouse L cells under the control of the mouse MT-I promoter. The secreted hGH in the medium was steadily accumulated during at least 10 days after transfection. When the MT-I promoter was induced by cadmium, the level of hGH was increased by several folds. Our results demonstrate that the plasmid vector containing the structural portion of the growth hormone gene can be used in transient expression systems for regulation studies.

LPS로 유발된 만성폐쇄성폐질환에 대한 생맥청폐음(生脈淸肺飮)의 영향

김용,양수영,김민희,남궁욱,박양춘,Kim, Yong,Yang, Su-Young,Kim, Min-Hee,NamGung, Uk,Park, Yang-Chun 대한한방내과학회 2011 大韓韓方內科學會誌 Vol.32 No.2

Objectives : This study aimed to evaluate the effects of Saengmaekcheongpye-eum (SCE) on a LPS-induced COPD (chronic obstructive pulmonary disease) model. Materials and Methods : The extract of SCE was treated to A549 cells and and LPS-induced COPD mouse model. Then, various parameters such as cell-based cyto-protective activity and histopathological finding were analyzed. Results : SCE showed a protective effect on LPS-induced cytotoxicity in A549 cells. This effect was correlated with analysis for caspase 3 levels, elastin contents, protein levels of cyclin B1, Cdc2, and phospho-Erk1/2, and gene expression of TNF-${\alpha}$ and IL-$1{\beta}$ in A549 cells. SCE treatment also revealed a protective effect on LPS-induced lung injury in COPD mouse model. This effect was evidenced via histopathological findings including immunofluorescence stains against elastin and caspase 3, and protein levels of cyclin B1, Cdc2, and Erk1/2 in lung tissue. Conclusions : These data suggest that SCE has pharmaceutical properties on lung injury. This study thus provides scientific evidence for the efficacy of SCE for clinical application to patients with COPD.

척수 좌상 후 트레드밀 운동이 피질척수로의 재생 및 신경생리학적 기능회복에 미치는 영향

김종오(Jong-Oh Kim),남궁욱(Uk Namgung),서태범(Tae-Boom Seo),오명진(Myoung-Jin Oh),윤진환(Jin-Hwan Yoon) 한국생활환경학회 2009 한국생활환경학회지 Vol.16 No.2

Currently, physical exercise is recommended as a therapeutic intervention to functional recovery in incomplete spinal cord injury patients. In the present study, we examined the effects of treadmill exercise on regeneration of corticospinal tracts and neurohysiological functional recovery after contusive spinal cord injury in rats. Adult rats were randomly assigned to one of three groups; (1) normal group; (2) SCI(contusion spinal cord injury) + sedentary group; (3) SCI(contusion spinal cord injury) + TMT(treadmill training exercise) group. One week after surgery, the treadmill exercise rats were exposed to treadmill low intensity exercise for 8 weeks. Using immunofluorescenestaining and anterograde tracing techniques, we identified enhanced axonal sprouting and regrowth in the corticospinal tracts after contusive spinal cord injury in rats with treadmill exercise. We also carried out western blot analysis to investigate axonal growth-associated protein (GAP-43) and Cdc2 expression in spinal cord. The neural functional assessment was determined by the BBB test. The results of present study showed that treadmill exercise increased GAP-43 and Cdc2 expression. Also, the results of present study showed that treadmill exercise decreased cavity formation. The treadmill exercise facilitated nerve regeneration in vivo by improving cell proliferation and migration, axonal sprouting and regeneration, and astrocytes after contusive spinal cord injury. It also accelerated functional recovery of hindlimb. In conclusion, the results of the present study suggest that treadmill exercise facilitates nerve regeneration and functional recovery after contusive spinal cord injury.

Sprague-Dawley 랫드를 이용한 선폐정천탕의 단회투여독성시험

이응석,한종민,양수영,김민희,김승형,남궁욱,박양춘,Lee, Eung-Seok,Han, Jong-Min,Yang, Su-Young,Kim, Min-Hee,Kim, Seung-Hyung,NamGung, Uk,Park, Yang-Chun 대한한방내과학회 2012 大韓韓方內科學會誌 Vol.33 No.1

Objectives : This study aimed to evaluate the single oral dose toxicity of Seonpyejeongcheon-tang (SJT) in male and female Sprague-Dawley rats. Methods : In this single oral toxicity study, rats were orally administrated in a single dose of 0 or 5,000 mg/kg SJT. There were 7 rats in each group. After single administration, mortality, clinical signs, body weight changes and gross pathological findings were observed for 14 days. Organ weight, clinical chemistry and hematology were tested after 14 days. Results : There was no mortality or other clinical signs for 14 days. There were also no significant differences in body weight, organ weights, hematological and serum chemical parameters between the SJT and control groups. Conclusions : The results obtained in this study suggest that the 50% lethal dose of SJT is over 5,000 mg/kg, so this finding can be expected to provide scientific evidence for the safety of SJT.

운동이 F344계 임신쥐에서 골격근의 VAMP-2 및 GLUT-4 단백질 발현과 혈중 인슐린, 렙틴 농도에 미치는 영향

윤진환,이희혁,김종오,오명진,박성태,지용석,서태범,남궁욱,Yoon, Jin-Hwan,Lee, Hee-Hyuk,Kim, Jong-Oh,Oh, Myung-Jin,Park, Seong-Tae,Jee, Young-Seok,Seo, Tae-Beom,NamGung, Uk 한국생명과학회 2007 생명과학회지 Vol.17 No.6

본 연구는 임신횐쥐에서 운동이 혈중 인슐린과 렙틴 농도, 그리고 골격근의 GLUT-4와 VAMP-2단백질 발현에 변화를 조사하기 위하여 수행되었다. 본 연구에서 비임신 횐쥐에 비해 임신 횐쥐에서 혈중 인슐린 농도가 유의하게 증가되었고, 렙틴 농도는 유의하게 감소하는 것으로 나타났다. 임신횐쥐에게 달리기 운동을 적용한 결과 임신으로 증가된 인슐린 농도를 유의하게 감소시키면서 렙틴 농도의 감소를 억제시키는 것으로 나타났다. 골격근에서 GLUT-4와 VAMP-2 단백질 발현량을 조사한 결과 비임신 횐쥐에 비해 임신 횐쥐에서 이러한 단백질 발현이 유의하게 감소되었지만, 임신 중 달리기운동을 경험한 횐쥐에서 GLUT-4와 VAMP-2 발현 모두 유의하게 증가되는 것으로 나타났다. 이와 잘이 임신으로 인한 혈중 인슐린 및 렙틴의 변화가 골격근에서 당을 근조직으로 흡수하는 신호 전달 경로와 수송체의 발현 손상과 연관되어있음을 보여주는 것이다. 하지만 임신 횐쥐에서 달리기 운동은 인슐린과 렙틴 농도의 변화를 완화시킴으로써 골격근의GLUT-4와 VAMP-2발현을 유의하게 증가시키는 것으로 나타났다. The main objective of this study was to investigate the effects of exercise on serum insulin and leptin levels and GLUT-4 and VAMP-2 in skeletal muscles from the pregnant rats. F344 rats were randomly divided into four groups (n = 15 in each group): control group (CG), pregnant group (PG), pregnant running group (PR), and pregnant swimming group (PS). From the 15th day of pregnancy, animals in the running group were forced to run on treadmill for 30 min with a light intensity, while those in the swimming group were forced to swim in swimming pool for 10 min once a day for 6 consecutive days. The present result demonstrated that in pregnant rat group, serum insulin levels significantly in-creased and leptin levels significantly decreased. Skeletal GLUT-4 and VAMP-2 protein expression was significantly decreased in pregnant rats compared to non-pregnant rats. However, matenal running during gestational period alleviated pregnancy-induced changes in plasma insulin and leptin levels, and it significantly enhanced skeletal GLUT-4 and VAMP-2 protein expression. From those results, it can be suggested that running exercise during gestational period may improve glycemic control by up-regulating GLUT-4 and VAMP-2 protein expression.

좌골신경 손상 후 스트렙토조토신 유도성 당뇨병에 의한 축사재생의 지연 효과

황진연 ( Jinyeon Hwang ),김기중 ( Ki-joong Kim ),남궁욱 ( Uk Namgung ) 대전대학교 한의학연구소 2020 혜화의학회지 Vol.29 No.1

Objective: The goal of this study is to investigate whether peripheral axonal regeneration is affected by diabetes in experimental animals. Method: Sprague Dawely rat was injected with 45~50 mg/kg of streptozotocin (STZ) to generate an animal model of diabetes. Three months after STZ injection, sciatic nerve (2 cm length) was removed and the same length of nerve segments from STZ-injected animal or from control animal (CTL) was transplanted into STZ - injected animals (STZ - STZ and STZ - CTL respectively). Similarly, sciatic nerve segments from STZ - injected animal or from control animal were grafted into the control animals (CTL - STZ and CTL - CTL respectively). All animals were sacrificed 2 weeks after transplantation. Sciatic nerve sections were prepared and subjected to immunofluorescence staining analysis. Results: Immunofluorescence staining for NF - 200 showed that distal elongation of regenerating axons reached 40~80% of proximal neve in both CTL - STZ and CTL - CTL groups. However, distal elongation in both STZ - STZ and STZ - CTL groups were 20~60% of proximal nerve. Furthermore, measurement of axonal regeneration after immuno - staining with SCG10 showed that the scores of distal elongation relative to proximal nerve were 50~90% in CTL - CTL and CTL - STZ groups and 10 - 60% in STZ - CTL and STZ - STZ. Conclusions: Our data showed that the levels of axonal regeneration were not affected irrespective of whether they were from STZ- or CTL graft, but were greatly reduced when the nerves were transplanted into the STZ host.

Effects of Exercise on Axonal Regeneration and Growth-associated Protein (GAP­43) Expression Following Sciatic Nerve Injury in Rats

서태범,윤성진,김경태,윤재석,윤진환,박성태,한인선,남궁욱,Seo Tae-Beom,Yoon Sung-Jin,Kim Kyung-Tae,Yoon Jae-Suk,Yoon Jin-Hwan,Park Sung-Tae,Han In-Sun,Namgung Uk Korean Society of Life Science 2005 생명과학회지 Vol.15 No.3

Growth-associated protein (GAP-43)은 손상된 신경 재생 시 axonal growth를 확인할 수 있는 가장 널리 알려진 단백질이다. 본 연구의 목적은 좌골신경 손상을 준 쥐에 저강도 운동 수행 후 좌골신경에서 발현되는 GAP-43 단백질의 양을 정량적으로 분석하고, 손상된 부분으로부터 신경섬유의 재생 정도를 조직학적으로 분석함으로써 운동이 말초신경 손상을 가한 쥐에 미치는 영향을 분석하는 것이다. CAP-43의 정량적인 분석은 western blotting을 통해 확인하였고, 좌골신경의 조직학적인 분석은 immunofluorescence staining과 anterograde로 DiI 주입, 척수 내의 motor neuron은 retrograde로 DiI를 주입 한 후 형광현미경에서 관찰하였다. 본 연구에서는 좌골신경이 손상된 쥐에게 운동을 수행시킴으로써 CAP피3단백질이 비운동 그룹보다 더 많은 양이 발현되고, 손상된 부위에서 재생하는 신경섬유의 길이와 양이 더 많은 것을 확인할 수 있었다. 또한 운동을 수행한 그룹에서 DiI에 염색된 motor neuron의 수가 더 많이 증가하는 것을 관찰하였다. 따라서 이러한 결과들은 운동이 좌골신경이 손상된 쥐의 재생을 촉진시킬 것으로 생각된다. Physical activity can improve sensorimotor recovery after peripheral nerve injury. Growth-associated protein 43 (GAP-43) is highly correlated with neuronal development and axonal regeneration and present in large quantities in the axonal growth cone. Using immunofluorescene staining and anterograde and retorgrade techniques, we identified enhanced axonal regrowth in distal stump of the sciatic nerve 3-14 days after crush injury in rats with treadmill training. We also carried out western blot to investigate GAP-43 protein expression in injured sciatic nerve. GAP-43 protein levels were highly induced in the injured sciatic nerve 3, 7 and 14 days compared with sedentary group. Thus, the present data provide a new evidence that treadmill training promoted axonal re-growth after injury and increased GAP-43 protein levels in the regenerating nerve.

좌골신경섬유 재생시 Cdc2 kinase 매개성 슈반세포 활성화의 역할 규명

한인선 ( In Sun Han ),서태범 ( Tae Beom Seo ),김종오 ( Jong Oh Kim ),남궁욱 ( Uk Namgung ) 대전대학교 한의학연구소 2005 혜화의학회지 Vol.14 No.1

맥문동탕(麥門冬湯)에 대한 연구 논문 분석

김형원 ( Hyoung Won Kim ),양수영 ( Su Young Yang ),김민희 ( Min Hee Kim ),남궁욱 ( Uk Namgung ),박양춘 ( Yang Chun Park ) 대전대학교 한의학연구소 2011 혜화의학회지 Vol.19 No.2

Objectives: This study analyzed the contents of the research papers of Maekmundong-tang (MMDT). This study was conducted to help development of new clinical application and clinical studies for treating COPD with Oriental medicine. Materials and Methods: We inspected 26 theses and scrutinized their classification, objective diseases, study design, participants, methodological quality of clinical trial. Results: The following results were obtained in this study. 1. The studies of MMDT started in 1989 and have continuously increased, but it decreased recently. 2. The studies were mainly focused on experimental models rather than clinical studies. The topics of studies were mainly relaxation of airway contraction and anti-asthmatic effect. 3. MMDT was showed to have effects on chronic cough and asthma in these papers. Conclusion: MMDT is being used in respiratory disease. However, mechanism study should be conducted at experimental study and more clinical studies on the efficacy of MMDT for chronic respiratory disease are needed.

청신도담탕(淸神導痰湯)이 식이성(食餌性) 고지혈증 동물 병태에 미치는 영향

이원구 ( Won Gu Lee ),이용구 ( Yong Gu Lee ),남궁욱 ( Uk Namgung ),김동희 ( Dong Hee Kim ) 대전대학교 한의학연구소 2004 혜화의학회지 Vol.13 No.2