갑상선암 세포주에서 아데노바이러스-hNIS(Ad-hNIS) 유전자 이입에 의한 방사성 요오드 포획 증진 효과

박건구,진정선,이성진,박정윤,이희란,문대혁,안일민,장혜숙 대한내분비학회 2000 Endocrinology and metabolism Vol.15 No.4·5

운동성 산화 스트레스와 항산화비타민의 보충이 말초단핵세포의 NF-kB 활성에 미치는 영향

진영수,박건구,박준영,김미정,이왕록,김혜영,이한준,박은경 대한스포츠의학회 2000 대한스포츠의학회지 Vol.18 No.2

Reactive oxygen species(ROS) are implicated in the pathogenesis of a wide variety of human diseases. Numerous studies indicate that ROS may serve as common intracellular molecules that contribute to the process of nuclear factor(NF)κB activation in response to a diverse stimuli. In our laboratory, we have demonstrated tat antioxidants could reverse the decline of immune function caused by exercise-induced ROS. Furthermore, it in necessary to understand a mechanism underlying ROS-dependent disorder in biological system. Recent studies have been shown that several gene expression were regulated by oxidants, antioxidants and other determinants of the intracellular reduction-oxidation(redox) state. In this process, NF-κB have been shown to play a important role. The purpose of the present investigation was to evaluate the effect of exercise-induced oxidative stress and antioxidnt supplementation on NF-κB activation in peripheral mononuclear cells. Forty male SD rats(4 weeks old) were randomly divided into noraml diet group and antioxidants(ATO) supplement group, and then ATO groups were treated with antioxidants(VE: 2001U/kg, VC: 50mg/rat, β-carotene: 300mg/kg, vitamin B6: 250㎍/100g, selenomethionine: 0.1mg/kg) for 16 weeks. After 16 weeks breeding at each condition, each group divided into two groups: Control group(CR) fed generally, Exercise group(CE) fed generally and followed by acute exercise 16 weeks later. Antioxidant Antioxidants and Exercise group(AE) fed with antioxidants and followed by acute exercise. The activation of NF-κB binding activity was increased after exhaustive exercise bout in both group. In addition, pretreatment of ATO group with the antioxidants mixture lead to the inhibition of NF-κB binding activity. This results suggest that NF-κB activation should be further studies in response to a variety of exercise.

정보제공자와의 면담에 의해 진단된 뇌졸중후치매의 유병률과 상관인자에 관한 연구

배희준,이건세,김형수,김병건,구자성,권오현,박종무 대한치매학회 2004 Dementia and Neurocognitive Disorders Vol.3 No.2

Background and Objectives: With increasing age of population, stroke and dementia become greater health problems in Korea. However, there have been no studies on poststroke dementia in Korea. We intended to elucidate the frequency and clinical correlates of poststroke dementia in a hospital-based cohort. Methods: From July 2001 to July 2002, 372 patients were hospitalized to Eulji General Hospital within 7 days from onset with acute stroke or transient ischemic attack. Two hundred-eight patients (55.9%) were followed up and interviewed more than 1 year later (451 ±83.3 days). Dementia was diagnosed by Korean Dementia Screening Questionnaire (KDSQ) obtained by a direct interview. Based on Eulji Stroke Registry, demographic factors, risk factors for stroke, stroke characteristics, and stroke outcomes were gathered and examined. Results: Of 208 patients, poststroke dementia was detected in 36 patients (17.3%, 20 males, age=64.5±9.4 years). Poststroke dementia was associated with history of stroke, aphasia, modified Rankin Disability Scale (MRDS) at 3 month after stroke and when interviewed, and Barthel index at 6 months after stroke (p;0.05). Its correlation with diabetes mellitus and National Institute of Health Stroke Scale at discharge were marginally significant (0.05;p;0.1). Conclusions: Post-stroke dementia is common in Korean stroke patients. Its frequency is comparable to that in other countries. The correlates of poststroke dementia and their clinical meanings are demonstrated.

췌장이식

한덕종,김인구,김석구,박건춘,민병철 울산대학교 의과대학 1992 울산의대학술지 Vol.1 No.1

Primary candidate for pancreas transplantation is patients with type Ⅰ diabetes mellitus of juvenile oneset. At present the patients selected for transplantation often have end stage renal disease, far advanced vascular disease and diabetic neuropathy. In these days, the survival of pancreas transplantation for treatments of patients with type Ⅰ diabetes mellitus has been improved especially in simultaneous pancreas and kidney transplantation. Various innovations in immunosuppression, management of the pancreatic ductal exocrine secretion, recipient selection and HLA matching have been contributed to achieve a reasonable outcome in pancreas transplantation compared with other organ transplantation. Current improved graft survival has stimulated many centers in the world to start pancreas transplantation program. Recently we successfully performed the first case of simultaneous pancreas and kidney transplantation in Korea. After 5 cases of pancreas transplantation in our department in type Ⅰ DM, we think that it is the time to review the pancreas transplantation to give an insight of this new field to our colleagues.

Gene Analysis Technology: the New M illenium of Therapy and Diagnostics

Kun-Koo Park 대한약학회 2004 大韓藥學會 總會 및 學術大會 Vol.2004 No.1

이온화 방사선에 의한 TIMP1 , TIMP2 유전자 발현 측정

박건구(Kun-Koo Park),진정선(Jung Sun Jin),박기영(Ki Yong Park),이연희(Yun Hee Lee),김상윤(Sang Yoon Kim),노영주(Young Ju Noh),안승도(Seung Do Ahn),김종훈(Jong Hoon Kim),최은경(Eun Kyung Choi),장혜숙(Hyesook Chang) 대한방사선종양학회 2001 Radiation Oncology Journal Vol.19 No.2

목 적 :Tissue inhibitor of matrix metalloproteinase (TIMP)는 matrix metalloproteinase (MMP)에 작용하여 암세포의 침윤과 전이를 억제하고 염증, angiogenesis, fibros is에 중요한 역할을 한다. TIMP 유전자는 여러 cytokine 및 signal molecule에 의하여 조절되는 유전자이므로 방사선에 의한 TIMP의 발현을 측정하고 전사 조절 기전을 연구하고자 하였다. 대상 및 방법 :두경부암 환자의 병변에서 유도하여 확립한 두경부암 세포주를 이용하여 방사선에 의한 TIMP 유전자 발현을 측정하였다. 각 세포주의 방사선 민감도를 측정하고 transwell을 이용한 invasion assay로 전이성을 측정하였다. TIMP1, TIMP2 발현은 conditioned medium을 취해 ELISA assay로 측정하였다. 방사선조사는 2 Gy, 10 Gy군으로 나누어 관찰했고 조사 후 시간 간격은 24, 48시간이었다. MTT assay로 생존세포 수를 측정하여 방사선 세포치사로 인한 발현 변화를 보정하였다. hTIMP1 promoter region을 PCR하여 pGL2- basic luciferase reporter vector에 cloning하여 인간 두경부암 세포주에 이입하여 functional TIMP1 발현이 증가하는지 확인하였고 protein kinase C(PKC) activator인 PMA (phorbol 12-myristate 13- acetate)와 Ras에 의한 TIMP1 발현이 유도되는지 확인하였다. 결 과 :HN- 1, HN-2, HN-3, HN-5, HN-9 세포주의 D0는 각각 1.55 Gy, 1.8 Gy, 1.5 Gy, 1.55 Gy, 2.45 Gy 이었다. 각 세포주의 방사선조사 후 MTT assay에 의한 cell viability는 24, 48시간에서 2 Gy인 경우 모두 94% 이상 그리고 10 Gy에서는 73% 이상의 생존 세포를 확인하였다. TIMP1, TIMP2 단백의 basal 농도는 24시간 48시간에서 점점 증가하여 세포에서 계속 합성되어 분비되고 있음을 확인하였다. 2 Gy 조사 후 24시간에서 TIMP2는 HN- 1, HN-9 세포주에서 감소하였으나, 10 Gy 조사 후에는 두 세포주에서 모두 증가하여 방사선량에 따라 반응이 달랐고, 방사선조사 후 48시간에는 HN- 1세포주에서는 증가하나 HN-9 세포주에서는 감소하여 세포주에 따라 반응이 달랐다. 그러나 방사선에 의한 TIMP1 발현 변화는 미미하였다. TIMP1 reporter gene을 인간 두경부암 세포주에 transfection하고 PMA (100 ng/ml)을 가한 경우 HN- 1세포주에서는 유의하게 증가하고 HN-9 세포주에서는 감소하였다. Ras 발현 벡터와 co- transfection한 경우 TIMP1 promoter가 활성화 되었다. 결 론 :모두 두경부 암에서 유래된 세포주 이지만 방사선에 의한 TIMP의 발현 및 전사조절 기전은 세포주 마다 차이가 있었고 이온화 방사선의 용량에 따라서, 방사선조사 후의 시간 경과에 따라서도 TIMP 발현에 차이가 있었 다. 이 결과는 TIMP의 전사 및 발현이 여러 종류의 signal molecule에 의하여 영향을 받고, 이 signal molecule들이 각 세포주 마다 다르기 때문으로 사료된다. Purpose : Express ion of TIMP, intrins ic inhibitor of MMP, is regulated by s ignal transduction in response to genotoxins and is likely to be an important step in metastas is , angiogenes is and wound healing after ionizing radiation. Therefore, we studied radiation mediated TIMP express ion and its mechanism in head and neck cancer cell lines . Materials and Me thods : Human head and neck ca ncer cell lines established at Asan Medical Center were used and radiosens itivity (D0), radiation cytotoxicity and metastatic potential were measured by clonogenic assay, MTT assay and invas ion assay, respectively. The conditioned medium was prepared at 24 hours and 48 hours after 2 Gy and 10 Gy irradiation and express ion of TIMP protein was measured by Elisa assay with specific antibodies against human TIMP. hTIMP1 promotor region was cloned and TIMP1 luciferase reporter vector was constructed. The reporter vector was transfected to AMC- HN- 1 and-HN-9 cells with or without express ion vector Ras , then the cells were exposed to radiation or PMA, PKC activator. EMSA was performed with oligonucleotide (- 59/- 53 element and SP1) of TIMP1 promotor. Results : D0 of HN- 1, - 2, - 3, - 5 and - 9 cell lines were 1.55 Gy, 1.8 Gy, 1.5 Gt, 1.55 Gy and 2.45 Gy respectively. MTT assay confirmed cell viability, over 94% at 24hrs , 48hrs after 2 Gy irradiation and over 73% after 10 Gy irradiation. Elisa assay confirmed that cells secreted TIMP1, 2 proteins continuous ly. After 2 Gy irradiation, TIMP2 secretion was decreased at 24hrs in HN- 1 and HN- 9 cell lines but after 10 Gy irradiation, it was increased in all cell lines . At 48hrs after irradiation, it was increased in HN- 1 but decreased in HN-9 cells . But the cha nge in TIMP secretion by RT was mild. The transcription of TIMP1 gene in HN- 1 was induced by PMA but in HN-9 cell lines , it was suppressed. Wild type Ras induced the TIMP- 1 transcription by 20 fold and 4 fold in HN- 1 and HN- 9 respectively. The binding activity to - 59/- 53, AP1 motif was increased by RT, but not to SP1 motif in both cell lines . Conclusions :We observed the difference of expresson and activity of TIMPs between radiosens itive and radiores istant cell line and the different s ignal transduction pathway between in these cell lines may contribute the different radiosens itivity. Further research to investigate the radiation response and its s ignal pathway of TIMPs is needed.

SHR(Spontaneously Hypertensive Rat)를 이용한 송엽, 익모초 추출물의 항고혈압 작용

박건구 ( Kun Koo Park ),류재원 ( Je Won Ryu ),최은경 ( Eun Kyung Choi ),노환성 ( Hwan Seong Ro ) 한국응용약물학회 2000 Biomolecules & Therapeutics(구 응용약물학회지) Vol.8 No.1

[S4-2] Gene Expression Changes Induced by Taurine in Cultured Human Hepatoma Cells, HepG2

Sung-Hee Park,Haemi Lee,Kun Koo Park,Ha Won Kim,Taesun Park 한국식품영양과학회 2005 한국식품영양과학회 학술대회발표집 Vol.2005 No.10

Gerbil 삼차신경절에서 허혈-재관류 후 calbindin D-28k와 calretinin 면역반응의 경시적 변화

황인구(In Koo Hwang),박정훈(Jeong-Hoon Park),최우제(Woo-Je Choi),박노진(Noh-Jin Park),오해수(Hae-Soo Oh),안성진(Sung Jin An),박승국(Seung-Kook Park),윤대근(Dae-Kun Yoon),이원학(Won-Hak Lee),강태천(Tae-Cheon Kang),원무호(Moo Ho Won) 대한해부학회 2002 Anatomy & Cell Biology Vol.35 No.5

허혈에 대한 연구는 중추신경계통에서 대부분을 차지하고 있다. 그러나 허혈은 중추신경 뿐 아니라, 말초신경에도 많은 영향을 미칠 것으로 생각된다. 특히 머리 부분에 있는 대부분 장기들의 일반감각을 담당하는 삼차신경절은 매우 중요할 것으로 생각한다. 신경전달물질의 분비와 감각의 조절 등에 중요한 칼슘결합단백질은 세포질내 칼슘을 조절하여 감각의 전달에 매우 중요하다. 따라서 본 연구는 Mongolian gerbil의 삼차신경절에서 인위적으로 허혈-재관류를 유발하여 calbindin D-28k (CB)와 calretinin (CR)의 경시적인 변화와 그 의미를 연구해 보고자 하였다. 정상군에서 CB과 CR 면역반응은 큰 크기, 중간 크기, 작은 크기의 신경세포에서 관찰되었다. 허혈-재관류 후 12시간까 지는 면역반응세포 수의 변화에 큰 차이를 관찰할 수 없었다. 허혈-재관류 후 1일 경에는 CB과 CR을 함유한 큰 신경세 포의 수가 급격하게 증가하였으며, 그 이후부터 CB 면역반응세포는 감소하였고, CR 면역반응세포는 허혈-재관류 후 4일 경에 중간 크기 신경세포가 정상군보다 3배 가량 증가하였다. 이상의 연구 결과는 허혈-재관류 후 1일 경에 큰 신경세포에서 CB와 CR 면역반응의 증가가 있었는데, 큰 신경세포들 은 A fibers를 함유하고 있으므로, 이 시기에 CB와 CR이 기계적 자극의 전도에 영향을 줄 것으로 생각이 된다. 또한 허 혈-재관류 후 4일 이후에 중간 크기의 신경세포에서 CR 면역반응 세포의 증가는 이들이 Aδ 또는 C fiber를 함유하고 있 으므로, 이 시기에는 통증이나 온도 감각에 CR이 관여할 것으로 생각된다. Many researches have focused upon temporal changes of neurotransmitters and/or neuromodulators in the central nervous system after ischemic insult. In sensory neurons, the spatial and temporal alterations of neurotransmitters have been little studied. Calbindin D-28k (CB) and calretinin (CR) have been suggested to play a role in the transmission of neurotransmitters. Therefore, in the present study we investigated the chronological alteration of CB and CR immunoreactivity in the trigeminal ganglion cells of the Mongolian gerbil after ischemic insult. In the sham operated group, CB and CR immunoreactivities were found in small-, medium- and large-sized neurons. One and two days after ischemia-reperfusion, small and large-sized CB immunoreactive neurons increased significantly. Thereafter, number of the CB immunoreactive neurons decreased markedly. Furthermore, five days after ischemia-reperfusion, CB immunoreactivity was detected in a few neurons, and its immunoreactivity was also very weak in the cytoplasm. Number of the large-sized CR immunoreactive neurons increased significantly one day after ischemia-reperfusion. Thereafter, the number of the large-sized CR immunoreactive neurons decreased. Especially, the number of the medium-sized CR immunoreactive neurons increased dramatically 4 days after ischemia-reperfusion. These results suggest that an increase of CB and CR may play an important role in modulating the mechanoception 1 day after ischemia-reperfusion, because the immunoreactivities increased in large-sized neurons which have the myenlinated A fibers. These results also suggest that significant increase of CR expression in medium-sized neurons 4 and 5 days after ischemia-reperfusion may provoke CR in modulating the nociception or thermoception because the medium-sized neurons which have the myenlinated Aδ or C-fibers.

인체 간암세포주(Hep 3B)의 증식에 미치는 amiloride의 억제효과

신채희,박기룡,최병주,서수홍,김성훈,박무인,박선자,정근옥,박건영,구자영 고신대학교 의학부 2002 高神大學校 醫學部 論文集 Vol.17 No.1

Background/Objective Cytoplasmic alkalinization induced by activation of the Na+/H+antiporter which is stimulated upon the addition of growth-promting agents, such as insulin, epidermal growth factor, phorbol ester, plays an essential role in the initiation on cell proliferation. In the present study the effects of amiloride, a specific and reversible inhibitor of Na+/H+exchanger, on the growth of human hepatocellular carcinoma cell line, Hep 3B were examined and the effects of 5-fluorouracil (5-FU) combined with amiloride were also studied to determine the role of amiloride in the treatment of hepatocellular carcinoma. Cell cycle analysis was done to examine the mechanisms for the inhibitory effects of amiloride. Methods The growth of Hep 3B cells were examined by counting cell number on two and four days treatment with 1μM, 10μM, 20μM, 40μM, 80μM, 160μM, amiloride, and 0.1㎍/㎖, 0.3㎍/㎖ 5-FU, after plating Hep 3B cells into 35-mm^(2) plastic dishes at a density of 20×10^(4) cells/dish. The reversibility of the effects of amiloride was examined on two days to eight days treatment with 20μM amiloride after seeding 4×10^(4) cells/dish Cell cycle analysis was done on the cells after four days treatment with 20μM amiloride. Results Amiloride significantly inhibited the growth of Hep 3B in a dose-dependent fashion (p<0.05). The inhibitary effect of amilride on the growth of Hep 3B cells was firstly shown at the concentration of 1μM, which is not so higher than the concentration of 0.1-0.2μM attainable by administration of usual dose of amiloride (5∼10㎎). Forty-eight percent inhibition of growth was found at an amiloride concentration of 20μM and 92% inhibition of growth was found at an amiloride concentration of 160μM after 4days treatment. The removal of amiloride by a media change after 48 hours treatment lead to significantly more growth than amiloride treated group (p<0.05). Amiloride combined with 5-FU significantly inhibited the growth of Hep 3B in a dose-dependent fashion compared to an amiloride or a 5-FU of cells in G0-G1 phase, S phase and G2-M phase was 57.9%, 20.8%, 21.3%, respectively in the amiloride group (20μM) and 38.9%, 20.8%, 40.3% in the control group, showing much higher G1 fraction in amiloride group compared to control group. Conclusions Amiloride significantly inhibited the growth of Hep 3B in dose-dependent fashion, which may be reversible. The reversibility of growth inhibition suggests that amiloride is not a non-specific cytotoxin for Hep 3B cells. Because the lowest inhibitory concentration of amiloride for the growth of Hep 3B cells in this study was 1μM, which is not so higher than the concentration of 0.1∼0.2 μM attainable by administration of usual dose of amiloride(5∼10㎎). amiloride or its analogues may be used alone or in conjunction with other modalities of therapy of hepatocellular carcinoma, for example, hepatic arterial chemoembolization of radiofrequency interstitial thermal ablation therapy. Since Hep 3B cell transition through the G1 phase was inhibited by amailoride, the inhibitory effects of amiloride on the growth of Hep 3B may be mediated in part by blocking G1-S transition. Further study is needed to clarify the effects of more potent analogues of amiloride on the growth of human hepatocellular carcinoma.