녹차 등에 의한 Peroxisome Proliferator-activated Receptor Transactivation의 변화 연구

이국경,조현영,주종호,김세일,황명실,염영나,강호일,김옥희 식품의약품안전청 2000 식품의약품안전청 연보 Vol.4 No.-

녹차는 동서양에서 오래 전부터 즐겄 마션오던 차이다. 암 예방효과가 있다고 여겨진 녹차가 랫드에서 과산화소체의 수와 관련효소인 paluitoyl CoA oxydase(PCoA)의 활성을 증가시킨다고 최근에 보고죄었다. 본 연군에서는 녹차에 의한 과산화소체의 증가간 peroxisome pro넓craters-activated receptor a (PPARO)의 활성을 통하여 일어나는 가를 알아보기 위하여 transient transfection assay법을 이용하였다. Cos-1 세포에 PP·ARu를 transfection 시킨 후 녹차, 홍차, 우롱차의 추출액과 녹차의 주성분인 epigallocatechin gallate (EGCG), epigallate catechin (EGC), epicatechin gallate (ECG),epicatechin (EC), gallic acid를 투여하였다. 또한 강력한 과산화소체 증가물질인 Wy-14,643과의 상호작용을 관찰하기 위하여 Wy-씨,643에 차 추출액 또는 성분들을 병용투여하였다. 녹차와 홍차 추출액과 EGCG(lOoM), EGC는 대조판에 비하여 PPARa의 활성을 1.5-2배 정도 증가시켰으며 차의 종류별로는 륵차>흥차>우롱차』둥글레차의 순으로 PPARC를 활성시켰다. 병용투여의 경우, 녹차는 Wy-14,643에 의한 PPARa의 활썽을 4-6배정도까지 강력하게 증가시켰다. 따라서 녹차는 PPARa에 직접적으로 작용하여 과산화소첸의 수를 증가시키는 것 이외에, 알려지지 않은 기전을 통하여 PPARa 의 활성조절에 관여하는 것으로 여겨진다. 또한 PPAaa의 찰성이 중간발효산물인 우롱차의 경우에는 낮은 반면 완전발효산물인 흥차에서 다시 높아지는 결과는 EGCG 이외에도 PPARa의 활성에 영향을 미치는 또 다른 물질이 존재함을 쩨시하고 있다. Tea is a number of peroxisomesnITnlber of oeroxisomespopular beverage. Recently it was reported that green tea increases the in rats. In this stfdy, to know that the rrlechanism to increase theanri activities of neroxilomal pnfvmpl il pxprrrf ihrourh activation of peroxisome proliferator-activated receptor u(PPARa), Cell trallsient transfection studies were carried out to investigate the interactions of tea extracts and major tea components with PPARa, cloned frorrl moust. Green tea and black tea extract, and epigallocatechlh gallate (ECCG), amajor component of fresh green tea leaves, slightly increased 1.5-2 times of the activatioR ofPPARC compared wiff DMEitf alone treatment. Especially the effective doses of tea extractsand EGCG were lower than doses necessary to EGCG-induced cell death and inhibition ofmetastasis by inhibiting urokinase activity In ether reports. Also at these concentrations, gieentea and bfack tea extracts enhanced the activation of PPARa induced by Wy-14,643(20 r M) ina dose dependent Tnanner. But even though oolong tea is semi-fermented during themanufacturing process, oolong tea extract showed less active than black tea_ These resvltsdemonshate that green tea possesses an unknown potent regulatory role in activation of PPARO byperoxisoine protiferators and some chemicals in addtion to EGCG may have role in thisregulation and exist iii the futl-fermented balck tea.

Peroxisome proliferator에 의해 발현 억제되는 유전자의 검색

이국경,강호일,김세일,염영나,정자영,황명실,김옥희 식품의약품안전청 1998 식품의약품안전청 연보 Vol.2 No.-

설치류예서 peroxisome pl·oliferator(PPs)의 장기투여는 간세포의 증식을 일으키며 이에 따른 간장 무게의 증가를 유발한다. 이후 간장 무게의 증가된 상태가 계속 유지되다가 수개월 후에 간암을 발생한다. 본 실험은 PPs에 의하여 간장 무게가 증가된 간세포의 상태를 유전자 발현순준에서 이해하기 위하여 dfferential display-reverse tran scrlption-polymerase chairl reaction(DD-HT-PCR) 기법을 이용하먼 발현이 감소된 유전자를 검색하였다. 이에 82개의 발현 감소가 예측되는 CBNA 절편을 분리하였고 reverse northerB blot으로 9종윽 유전자가 발현 감소됨을 확인할 수 있었다. 9종의 선별된 CDNA 절편의 염기서열을 확인한 곁과 2종은 transferrin과 of-inhibifur 111에 대해 각각 100%읜 동질성을 갖는 CDNA 절편이었으며, 또 다른 2종의 CDNA 절펀은 long Inter spersed repetitive nuclear element(LINE)와 98%의 동질성을 보였단. 나떠지 5종윽 CDNA 절편은 매우 낮은 농질성을 보이거나 꿀색이 되지 않았다. Transferrin의 감소는 peroxisome pro?iferator actiuated receptor(PPAR)가 transferrin의 transcription factor인 HNF-4(hepatocyte nuclear factor-4)와 경쟁적인 작용함에 기인한다.본 실험에서 PPs에 의해 발현 감소되는 것으로 처음 밝혀진 α1-inhibitor III는 종양세포에서 발현이 감소하는 유전자로 알려져 잇으며 또한 역시 PPs에 의해 발현감소되는 retrotransposon인 LINE과 아직 동정되지 않은 5종의 유전자들의 발현 감소는 PPs의 작용범위가 알려진 범위보다 넓을수 있음을 제시하고 있다. 따라서 PPs는 현재까진 알려진 transcription factor외에도 다른 transcription factor와 경쟁적으로 aL울괴거나 또는 다른 경로를 통하여 단백질의 발현에 관여함으로써 간세포의 증식 또는 대사에 영향을 끼칠 수 있을 것으로 여겨진다. Peroxisome proliferators (PPs) increase the liver weight. maintain the increased weight of liver. and eventuallf influce liver tumor in rodents. To understand the hyferplastic status of liver in-duced by long-terrE treatuent of PPs, the techaique ef differential display-reverse transcription-poly-morass chain reaction (D3)-RT-PCR ) was rerformed to detect the transcriptionally down-regulatedgenes that would be relateil te the homeostasis or altered function of rat liver treated with PPs. To iso-late HNA from rat liver maintaining increased weight, the rats were given 0.5% clofibrate-containingdiet for 3 weeks. Using DD~RT-PCR teehnique,82 suspected CDNA fragnlents shewing transcriptionallydown-regulated patterns were isolated. Only 9 CDNA fragments aiuoBg these fragraents were decreasedtranscriftionally by 4 other PPs (Wy-14,643, di(2-ethylhexyl) phthalate, di(2-ethylhexyl) adipate, andperchloroethylene). Two CHNA fragments have 100% similarity wi·th transferrin and α1-inhibitor lIt,respectively. Two other CBNA fragmeBts have 98% sirailarity with long interspersed repetitive nuclearelements (LINE). The expression of transferrin was already knowB to be regulated by HNF-4(hepatecyte nuclear factor-4) and decreased by competitive action of peroxisome proliferator-activated.scepter :fPAR) . But the t.anscriptionally down-.egulatioa of of-inhibitor lIT by PPs was firstlyshown in-tliis study. The transcriptional reductions of a nlunber of LINE interspersed within raammali-an genome and the unidentified genes imply that the scope of genomic targets by PPs may be more enor-mous than what we have krㄴown. Therefore these results svggest that PPs may have an influence on cetlcycle or homeostasis througt competitive action against transcription factors or through other pathway.

생체 외 제대혈 배양에서 거대핵세포 조혈에 대한 Interleukin-11 (IL-11)의 효과

이국경,김찬규,이남수,김숙자,정희정,이규택,박성규,백승호,원종호,홍대식,박희숙,Lee, Kuk-Kyung,Kim, Chan-Kyu,Lee, Nam-Su,Kim, Sook-Ja,Cheong, Hee-Jeong,Lee, Kyu-Tack,Park, Sung-Kyu,Baick, Seung-Ho,Won, Jong-Ho,Hong, Dae-Sik,Park, Hee-So 대한면역학회 2003 Immune Network Vol.3 No.1

Background: The megakaryopoiesis and platelet production is regulated by several hematopoietc factors such as thrombopoietin (TPO), interleukin-11 (IL-11) and interleukin- 3 (IL-3). IL-11 is a potent stimulator of megakaryopoiesis in vivo, and acts primarily as a megakaryocyte maturation factor in vitro and it can act synergistically with IL-3 and TPO. We performed this study to investigate the effects of recombinant human IL-11 (rhIL-11) with other hematopoietic factors on megakaryocyte colony formation in vitro. Methods: CD34+ cells were separated from umbilical cord blood and megakaryocyte colonies using MegaCult Assay Kit were cultured with rhIL-11, recombinant human IL-3 (rhIL-3), and recombinant human TPO (rhTPO) for 7 and 14 days. The number and percentage of CD34+ and CD41a+ cells were determined by flowcytometry. Results: The number of CD41a+ cells were $0.54{\pm}0.05{\times}10^4$ (rhIL-11 100 ng/ml), $5.32{\pm}0.23{\times}10^4$ (rhIL-3 100 ng/ml), and $8.76{\pm}0.15{\times}10^4$ (rhTPO 50 ng/ml) of total expanded cells during the culture of the purified CD34+ cells in liquid phase for 7 days. The number of CD41a+ cells were increased to $7.47{\pm}0.69{\times}10^4$ (rhIL-3+ rhIL-11), $11.92{\pm}0.19{\times}10^4$ (rhTPO+rhIL-11) of total expanded cells, respectively, during the culture of the purified CD34+ cells in liquid phase for 7 days in the presence of rhIL-11 (100 ng/ml). When the purified CD34+ cells were cultured in semisolid mediaincluding various concentration of rhIL-11, the megakaryocyte colonies were not formed. When the purified CD34+ cells were cultured with rhIL-11 and rhTPO or with rhIL-11 and rhIL-3, the number of megakaryocyte colonies were increased compared with rhTPO or rhIL-3 alone. Conclusion: These results indicate that IL-11 exerts a potent proliferative activity to colony forming unit-megakaryocyte from human umbilical cord blood, and it acts with other hematopoietic factors synergistically.

비혈연간 동종 골수이식술 후 발생한 혈전성 미세혈관병증 1례

이국경,배상병,김주성,이준혁,이유경,이규택,박성규,원종호,백승호,진소영,홍대식,박희숙 대한조혈모세포이식학회 2000 대한조혈모세포이식학회지 Vol.5 No.2

혈전성 미세혈관병증은 골수이식 후 발생하는 중한 합병증 중의 하나이다. 저자들은 22세의 급성 림프구성 백혈병(ALL, L3)을 진단 후 비혈연간 동종 골수이식을 받은 환자에서 발생한 혈전성 미세혈관병증을 경험하였다. 그는 비혈연간 동종골수이식술을 받았으며 이식의 전처치로 busulfan, cyclophosphamide, 전신방사선조사를 받았고 이식편대숙주반응을 막기 위하여 CsA와 methotrexate을 투여받았다. 이식 후 28일째 말초혈액도말검사상 Burr cell, schistocyte, tear drop cell이 관찰되었으며, 이식 후 31일째 혈변소견이 보여 S결장 내시경상 장관 이식편대 숙주질환 소견이 보였고 혈청검사상 거대세포바이러스가 양성소견을 보여 steroid와 gancyclovir를 투여받았고 2주 후에 장관 이식편대 숙주반응이 호전되었다. 이식 후 68일째 임상적으로 혈전성 미세혈관병증이 발생되었고 Cr 수치가 증가되어 TPE와 혈액투석을 격일로 시행하였으나 악화되었다. 이식 후 82일째 다발성 출혈로 사망하였다. Thrombotic microangiopathy (TMA) is one of the serious complications of bone marrow transplantation (BMT). We experienced a 22-year-old male with acute lymphoblastic leukemia (ALL, L3) who developed post BMT-TMA. He received pretransplant conditioning chemotherapy with busulfan, cyclophosphamide, and total body irradiation (TBI). He received cyclosporine (CsA) and methotrexate (MTX) for graft versus host disease (GVHD) prophylaxis. Peripheral blood smear revealed burr cells, schistocytes and tear drop cells on post BMT day 28. Hematochezia was developed on post BMT day 31. Biopsy specimen of sigmoid colon revealed grade III to IV acute GVHD and cytomegalovirus (CMV) culture was positive in blood on post BMT day 43. He received steroid pulse therapy and gancyclovir so that improved after 2 weeks. TMA was developed clinically on post BMT day 68 and creatine (Cr) was increased. In spite of undergoing plasma exchange with fresh frozen plasma and hemodialysis every other day, TMA progressed. He died due to multiple hemorrhage on post BMT day 86.

Diethylnitrosamine에 의한 랫드 간세포 Clone 9의 In vitro Transformation 특성

이국경,정자영,김옥희 한국수의공중보건학회 1999 예방수의학회지 Vol.23 No.1

포스타 전시 발표 : 소화기 ; 보존적 요법만으로 치유된 식도 점막하종양의 내시경적 절개핵적출술후 발생한 식도 천공

이국경,김영태,김진오,조주영,이문성,심찬섭 대한내과학회 1998 대한내과학회 추계학술대회 Vol.1998 No.1

물질의 약리기전 및 독성기전관리를 위한 데이터베이스

이국경,양원형,박원근,송태웅,허창복,박전홍,홍용원,김송학,오찬우 濟州大學校亞熱帶農業硏究所 2004 아열대농업생명과학연구지 Vol.20 No.1

Managing, processing, and expressing the data, the database system makes users save the time and labors required to look up and organize the useful data as well as helps to create some new information and ideas. However in Korean situation, our the history of research is not too long and the resources including database systems is not enough. Because other previous database svstems manage the data in a form of text file. I t is not easy to obtain a useful analyzed results at once. The goals of this project are to extract experimental data from the research on endocrine disruptors. to manage the data as a code form, to process the data, to create the useful analyzed results, to help toxicological researchers. and eventually to enhance the availability of raw data. Before the systems design, we constructed the code tables from the reports and executed a pilot test. Depending upon the result of pilot test, we determined the factors to be extracted as data set from reports. constructed database tables, and designed the software to manage the data-input and to search and analyze data, Data-input module help users easily to analyze the report in a paper form and to input the data. Developed in the web-based form, data-search module is able to give an access to public as well a s researchers. Also depending on the data saved in a code form. data-search module can provide much useful information and have a potentiation to be developed as more powerful data-analyzing system.

내분비계 장애물질 연구를 위한 데이터베이스

이국경(Kookkyung Lee) 한국실험동물학회 2004 Laboratory Animal Research Vol.20 No.1

Managing, processing and expressing data, the database system enables users to save their time and labor required for looking up and organizing useful data, as well as it helps to create some new information and ideas. For Korea, however, in the area of toxicological research, the history of toxicology hasn't been so lengthy and resources like the database system are not adequate. Because other previous database systems have managed the data in the form of a text file, it is not easy to quickly obtain a useful analyzed result. The goals of this project are to extract experimental data from the research on endocrine disruptors, to manage the data into a code form, to process the data, to create useful analyzed results, to help toxicological researchers, and to eventually facilitate the availability of the raw data. To achieve these goals, we first constructed the code tables from the reports, and then a pilot test was conducted. Depending upon the result of the pilot test, we determined the factors to be extracted as the data set from the reports. We then constructed the database tables and designed the software to manage the data-input. The data-input module helps users easily analyze the report in the paper form and also, to input the data. Developed in the web-based form, the data-search module is able to give access to the public, as well as to toxicological researchers. Because as the data is saved in a code form, the data-search module can provide much useful information and it has the potential to be a very powerful data-analyzing system.

3-chloro-1,2-propanediol(3-MCPD)의 독성학적 고찰

이국경(Kookkyung Lee) 한국실험동물학회 2004 Laboratory Animal Research Vol.20 No.1

3-monochloropropane-1,2-diol (3-MCPD) is a member of a group of compounds known collectively as chloropropanols, a group that include the genotoxic animal carcinogen, 1,3-dichloropropan-2-ol (1,3-DCP). 3-MCPD has been detected as a contaminant of several foods, food ingredients, including acid-hydrolysed vegetable protein, malts and soy sauces. Also 3-MCPD is a contaminant of polyamine flocculants used in the production of drinking water. It has given positive results in in vitro mutagenicity studies but negative results in in vivo mutagenicity test. 3-MCPD also itself is an animal carcinogen producing tumors at various sites in male F344 rats (mammary tissue, testes and preputial gland) and renal tubular adenomas and carcinoma in both sexes of F344 rats. However, except for the nephrotoxicity and antifertility in testes, renal tumor and mammary gland tumor could be promoted by chronic progressive nephropathy and spontaneous testicular tumor due to species-specificity of F344 rats, respectively. In addition, 3-MCPD has shown some effect on hormonal balance in F344 male rats. These results suggest that the additional test may be necessary to confirm the carcinogenicity and the possibility to be an endocrine disruptor.

플리스틱 가소제인 DIISOCECYL PHTHALATE 의 독성학적 고찰

이국경(Kook Kyung Lee),주종호(Jong Ho Joo),김세일(Se Yl Kim),제정환(Jeong Hwan Che),남기택(Ki Taek Nam),강진석(Jin Seok Kang),안병우(Byeong Woo Ahn),홍충만(Choong Man Hong),임국환(Kook Hwan Rhim),양기화(Ki Hwa Yang),장동덕(Dong Deuk 한국수의병리학회 2000 한국수의병리학회지 Vol.4 No.2