Percoll®의 비연속 밀도구배에 의한 Lymphocyte subpopulation의 분리

박재경 ( Jai Kyung Park ),최영주 ( Young Joo Choi ),김영일 ( Young Ii Kim ),이광길 ( Kwang Gil Lee ),김재만 ( Jae Man Kim ),남상윤 ( Sang Yun Nam ) 대한임상검사과학회 1988 대한임상검사과학회지(KJCLS) Vol.20 No.1

각질형성세포에서 HLA - DR 항원의 발현이 IL - 1 생산에 미치는 영향에 관한 연구

심우영,박재경,허충림 ( Woo Young Sim,Jai Kyung Park,Choong Rim Haw ) 대한피부과학회 1991 大韓皮膚科學會誌 Vol.29 No.6

N/A There is increasing evidence that keratinocytes may parcipitate in the inflammatory processes and immune response of skin. Keratinocytes can be induced to express functional immunocompetent molecules such as HLA-DR antigen. In additior, they secrete a wide range of infiammatory cytokines including IL-1. In order to better understand the immunologic role of keratinocyte, it is necessary to quantitate the amount of IL-1α produced by cultured human keratinocytes and evalute the effect of HLA-DR expression on the production of IL-1α by keratinocytes. In this study, we employed the radioimmunoassay to measure the amount of IL-1α produced by cultured human keratinocyte. Human keratinocytes were isolated from neonatal foreskins and grown in MCDB 153 medium. HLA-DR expression was induced by 30 U/㎖ of r-IFN-?. The supernatants were assayed for IL-1α. The results were as follows : 1. When culture medium was not replaced until 72 hours, cell numbers of kerantinocytes without treatment of r-IFN-? were 0.80±0.26×10^6, 1.16±0.21×10^6, 1.33±0.21×10^6 at 24, 48 and 72 hours, respectively. When culture medium was replaced every 48 hours, cell numbers of keratinocytes with treatment of r-IFN-? were 0.72±0.29×10^6, 1.06±0.26×10^6 and 1.06±0.25×10^6 at 24, 48 and 72 hours, respectively. Antiproliferative effect of r-IFN-? was significant at 48 and 72 hours(p=0.046, p=0.003). When culture medium was replaced every 48 hours, cell numbers of keratinocytes without treatment of r-IFN-? were 1.16±0.21×10^6, 1.69±0.27×10^6, 2.01±0.18×10^6 at 48, 96 and 144 hours, respectively. Cell numbers of keratinocytes with treatment of r-IFN-? were 1.06±0.26×10^6, 1.51±0.20×10^6 and 1.67±0.17×10^6 at 48, 96 and 144 hours, respectively. A?tiproliferative effect of r-IFN-? was significant at 48 and 144 hours(p=0.046, p=0.0001). 2. Relatively lower concentration of IFN-?(30 U/㎖) can induce HLA-DR expression on keratinocytes. The percent of HLA-DR positive keratinocytes were 42.15±4.87% by 48 hours. Then it decreased to 14.26±2.85% by 144 hours. 3. When culture medium was not replaced, the levels of IL-1α from supernatants of cultured keratinocytes were 1.513±0.774, 0.778±0.299 and 0.720±0.274 fmol/10^7cells at 24, 48 and 72 hours, respectively. The level is highest at 24 hours and then i? decreased. 4. When the percentage of HLA-DR positive keratinocytes were relatively high 42.15±4.87% and 38.59±3.74% at 48 and 72 hours, respectively, the levels of IL-1α were 1.009±0.320 and 1.002±0.301 fmol/10^7cells, respectively. They were significantly higher than r-IFN-? untreated groups, of which IL-1α levels were 0.778±0.299 and 0.720±0.274 fmol/10^7cells, respectively(p=0.059, p=0.007). When the percentage of HLA-DR positive keratinocytes were decreased, the differences of IL-1αlevel between two groups were not significant. In summary, the results indicate that the amount of IL-1α produced by keratinocytes are relatively low and keratinocytes may participate in the immune process of the skin, and that the immunologic role of HLA-DR^+ keratinocytes might be media?ed via an increased production of IL-1α from HLA-DR^+ keratinocytes.

UVB 반복조사가 배양 인체 멜라닌세포의 형태학적 변화와 증식 및 멜라닌화에 미치는 영향

김진환,박재경,허충림,이무형 ( Jin Hwan Kim,Jai Kyung Park,Choong Rim Haw,Mu Hyoung Lee ) 대한피부과학회 1994 대한피부과학회지 Vol.32 No.6

피부는 외부에 노출되어 있으므로 외부의 여러 물리적 또는 화학적 요소들에 의해 영향을 받아 변화하는데, 그중 일광은 항상 접하게 되는 필수적 요소의 하나이다. 일광이 피부에 미치는 영향에는 비타민 D의 합성 등의 인체에 이로운 작용외에도 급성반응으로 홍반이나 색소침착, 만성반응으로 나타나는 피부노화, 피부암의 유발 등은 인체에 유해하거나 미용적인 문제를 야기하게 한다. 이러한 피부변화는 표피세포 뿐만 아니라 진피세포들의 복합적인 작용으로 일어나게 된다. 그러나 표피나 진피를 구성하는 세포 각각의 독립적인 변화를 알아보기 위해서는 구성세포의 분리배양이 필요하다. 특히 멜라닌세포는 일광노출에 의해 멜라닌 합성이 증가됨으로써 피부의 색소침착에 중요한 역할을 담당할 뿐 아니라, 이 합성된 멜라닌이 각질형성세포로 이동함으로써 일광 등에 의한 피부의 손상을 방지하게 된다. 멜라닌세포의 성장과 멜라닌화에 영향을 미치는 요소에는 세포의 유전성 프로그램, 일광, 호르몬, 그밖의 멜라닌 대사에 영향을 주는 화학물질 등이 있다. 그중 일광노출에 의한 변화는 주로 자외선B(이하 UVB라고 약함)에 의해 일어나는데, 일반적으로 생체내 실험에서는 멜라닌 세포수가 증가되며 멜라닌화를 촉진하는 것으로 알려져 있다. 그러나 배양 멜라닌세포를 이용한 실험관내 실험에서는 보고자에 따라 다른결과를 보이는데, 이는 자외선의 조사방법, 조사량, 멜라닌세포의 배양방법, 종족이나 개체간의 멜라닌 세포의 특성의 차이 등으로 인해 나타난 것으로 생각된다. 특히 조사량과 함께 조사횟수 즉, 일회조사한 경우와 반복조사한 경우의 멜라닌세포수나 멜라닌 양은 다르게 나타난다. 저자는 일상생활에서의 일광노출과 유사한 적은 양의 UVB를 배양 인체 멜라닌세포에 반복 조사하여 멜라닌세포의 성장과 분화의 변화를 알아보았다. 또한 동일량의 UVB를 분할하여 반복조사한 경우와 일회조사한 경우에 어떤 차이가 있는지 알아봄으로써 UVB가 실험관내 실험에서 멜라닌세포의 성장과 분화에 미치는 영향을 규명하고자 본 실험을 시행하였다. Background:In the skin, the major stimulus for cutaneous pigineatation is ultraviolet radiation. The most important physiologic role of melanin is protection against harmful UV radiation to skin. It is known there are some differences in melanization between a single and multiple exposures of UB, in vivo. Little if known about the functions of the melanocyte alone in cutaneous pigmentation after ultraviolet exposure, because of the complexity of interactions in the whole epidermis Objective:To investigate the effects of multiple exposures at various dosages of UVB, and to compare the effect of UVB in multiple divided exposures ? a single exposure at the same total dosage of UVB on proliferation and melanization in cultured human melanocyte. Methods:Melanocytes were cultured by modified TIC medium The melanocter were exposed daily for three consecutive days to UVB at 2, 4, 8, and 16 mJ/㎠ and a single exposure at 24 mJ/㎠. The morphologic changes were examined by phase contrast microscopy. The melanocytes were counted by hemocytometer and melanin content;were assayed by spectro-photometer. Results:1. The effects of multiple UVB exposures: 1)The morphologic changes were as follows:With three time exposures at a dosage of 8 mJ/㎠, the melanocytes enlarged in size, and elongated their dendrites slightly;with three time exposures at a dosage of 16 mJ/㎠, enlargement in sized and elongation of dendrited were more significant. 2) With three time exposures at dosages of 2 and 4 mJ/㎠, the proliferation of melanocytes was stiumlated significantly(p<0.05). However, with three time exposures at dosages of i and 16 mJ/㎠ the proliferation was inhibited(p<0.05). 3) With three time exposures at dosages of 2 and 4 mJ/㎠, the melanin contents were decreased. However, with three time exposures at a dosage of 16 mJ/㎠, the melanin contents were highly increased(p<0.01). 2. The comparison between multiple divided exposures and a single exposure at the same toal dosage of UVB: 1) There were no morphologic differences of dendrities between with three time exposures at a dosage of 8 mJ/㎠ and with a single exposure at a dosage of 24 mJ/㎠. However enlarged melanocytes were more numerous with a single exposure. 2) The proliferation of melanocytes was more inhibited with:single exposure than with multiple divided exposures(p<0.05). 3) The melanin contents were more increased with a single exposure than with multiple divided exposures(p<0.05). Conclusion:With multiple exposures at lower dosages of UVB, the proliferation of melanocytes was stimulated, and melanixation was decreased. However, with multiple exposures at higher dosages of UVB, the proliferation was inhibited, and melanization was increased. At the same total dosage of UVB, the proliferation on was more inhibited, and the melanization was more increased with a single exposure than with multiple divided exposures.(Kor J Dermatol 1994;32(6):1035∼1045)

감마인터페론이 각질형성세포의 인터루킨 - 1 생산에 미치는 영향

오송,심우영,박재경,허충림 ( Song Oh,Woo Young Sim,Jai Kyung Park,Choong Rim Haw ) 대한피부과학회 1993 大韓皮膚科學會誌 Vol.31 No.1

각질형성세포에서 ICAM - 1 , HLA - DR 발현의 역동적 변화와 이들이 동종 말초혈액림프구와 혼합배양에 미치는 영향

국승현,심우영,박재경,허충림 ( Seung Hyun Kook,Woo Young Sim,Jai Kyung Park,Choong Rim Haw ) 대한피부과학회 1993 大韓皮膚科學會誌 Vol.31 No.5

행인 제제의 독성 및 항암효과의 자연살해세포 활성에 미치는 효과

심범상,최승훈,박재경,Shim, Bum-Sang,Choi, Seung-Hoon,Park, Jai-Kyung 대한암한의학회 2000 大韓癌韓醫學會誌 Vol.6 No.1

In this experiment, we examined the the effect of Laetrile Oil to the immune system and tried to disclose the anti-cancer mechanism of this. The result is listed as below. 1. After per-os, sub-cutaneous and into the peritoneal injection of Laetrile Oil to the SPF mice, the LD50 is above 5000mg/kg at even group. 2. Mean survival days of mice treated with laetril oil, after S-180 cells transplantation into the peritoneal cavity decreases 1.5 days(-6.8%) compared with the Mean survival days of the control group(22days.) 3. Effects of laetril oil on natural killer cell activity at Effector/Target Cell Ratio with 100:1, 50:1, 10:1 into methotrexate-pretreated mice is like this.: Compared with $29.22\pm12.7\%$ Cytotoxicity of the control group, sample group's Cytotoxicity had $38.83{\pm}12.5%$ of meaningful acrease. At 100:1 Effecr/Target Cell Ratio. At 50:1 Effector/Target Cell Ratio, control group has $20.02{\pm}9.6%$ Cytotoxiciy and sample group had $31.53{\pm}13.4%$ Cytotoxicity. At 25:1 Effector/Target Cell Ratio control group has $13.60{\pm}6.6%$ Cytotoxicity and sample group had $20.81{\pm}9.8%$ Cytotoxicity According to the above results, the Laetrile Oil represents nontotoxic to a SPF mice. non-effective to transplantable Sarcoma 180 tumors, and activaion in NK cell activity.

모유두세포의 배양에 관한 연구

심우영(Woo Young Sim),박재경(Jai Kyung Park),허충림(Choong Rim Haw) 대한피부과학회 1995 대한피부과학회지 Vol.33 No.1

Background : Dermal papilla cells, which are mesenchyral components of the hair bulb are considered to play a fundamental role in the inductior hair growth. Objective : We studied how to establish a method of cuku e of dermal papilla cells. Methods : With human scalp tissue, we cultured dermal pipilla cells and identified them under the double-contrat microscopy and observed the epassion of eactin. Results : In vitro they spread slowly, initially as a moayer, and eventually formed multilayered paralled anys. At the edges of expanding colonies the cells were large and flattened and showed a tendency to form clumps. They expressed a-actin which is thought to be a marker of papilla cells, typeIII, typeIV colagen and laminin. Conclusion : These results suggest that using the female papilla cells, specially differentiated fibroblast, may be useful for the study of hau growth. (Kor J Dermatol 1995; 33(1); 28-32)

배양된 인체 멜라닌 세포의 증식과 HLA - DR 항원 표현에 대한 재조합 감마 인터페론의 영향

어수동,이무형,엄주용,박재경,허충림 ( Soo Dong Ahu,Mu Hyoung Lee,Joo Yong Eom,Jai Kyung Park,Choong Rim Haw ) 대한피부과학회 1993 大韓皮膚科學會誌 Vol.31 No.3

Insulin-Like Growth Factor 1(IGF-1)이 멜라닌세포의 증식에 미치는 영향

조양훈(Yang Hoon Cho),박재경(Jai Kyung Park),이무형(Mu Hyoung Lee) 대한피부과학회 2000 대한피부과학회지 Vol.38 No.10

Background:Human growth hormone(hGH) plays a central role in linear bone growth and body metabolism. Its mitogenic effect in human tissues is mediated via direct and indirect actions. As proposed by the somatomedin hypothesis, many circulating GH-mediated effects are exerted indirectly and systemically via stimulation of hepatic synthesis of insulin-like growth factor 1(IGF-1). Given additional evidences for the expression of growth hormone receptor(GH-R) and IGF-1 receptor(IGF-1R) on many target tissues including keratinocytes, melanocytes, and fibroblasts, it is now evident that the GH can act via systemic IGF-1 secreted by the liver and locally produced IGF-1, as well as directly through the GH receptor.Objective:The purpose of this study was to investigate not only the effect of IGF-1 on the morphologic changes, proliferation, and melanization of cultured human melanocytes but also on its signal transduction pathway through the IGF-1R. Methods:Melanocytes were exposed to IGF-1 at 10, 25, 50, 75, 100ng/ml and we examined the changes of cell morphology, number of cells, [3H]-thymidine incorporation, MTS assay, and melanization according to the concentrations and exposure times of IGF-1. Also, the activity of p44/42 MAPK/ERK according to the various exposure times of IGF-1(25ng/ml) was examined using the Western blotting method to find out about the signal transdution pathway of IGF-1. Results : The results were as follows:1. There were no significant morphological changes of cells between the control and experimental groups according to the concentrations and exposure times of IGF-1. 2. The effects on melanocytes according to the concentrations of IGF-1 5 days after adding IGF-1 : 1) The number of cells, [3H]-thymidine incorporation, and MTS assay were significantly higher than those of control group in all experimental groups(p<0.05). 2) The melanin content showed an insignificant decrease in all experimental groups. (Korean J Dermatol 2000;38(10):1315~1324) 3) The melanocytes responded independent of the IGF-1 concentration in the assay of cell number, [3H]-thymidine incorporation and MTS. 3. The effects on melanocytes according to the exposure times(3 days, 5 days, 7 days) of IGF-1(25 ng/ml) : 1) The number of cells, [3H]-thymidine incorporation, and MTS assay increased as time went by, and was significantly higher than those of control group at all exposure times(p<0.05). 2) The melanin content decreased after exposure of IGF-1, especially that of 3 days exposure group showed a significant decrease(p<0.05). 4. The activities of p44/42 MAPK/ERK increased suddenly at 5 minutes with a peak at 60 minutes and then abruptly decreased at 120 minutes after adding IGF-1 Conclusion:In summary, this study demonstrates that IGF-1 has no effect on the morphology, but it does increase the proliferation and slightly decrease the melanization of cultured human melanocytes. In addition, it is suggested that IGF-1 plays a role in regulation of proliferation of melanocytes via the receptor PTK pathway with activation of p44/42 MAPK/ERK.

녹차추출물인 (-)-epigallocatechin-3-gallate와 polyphenol이 배양 인체 각질형성세포, 인체 유표피암세포, 섬유모세포, 멜라닌세포, 및 혈관 내피세포의 증식에 미치는 영향

최재영 ( Jae Young Choi ),김경달 ( Kyung Dal Kim ),박재경 ( Jai Kyoung Park ),김낙인 ( Nack In Kim ) 대한피부과학회 2002 대한피부과학회지 Vol.40 No.4

N/A It is known that skin diseases ar related to proliferation of cellular components. Green tea has m a v favorable biologic effects: anti-inflammatory, antibacterial. antiviral effects. lowering of plasma cholesterol and triglyadde levels, reduction of blood pressure and platelet aggregation, antimuragenic, and anticarcinogenic activities. The effects of green tea on various cells in skin has been reported in previous studies; For keratinocytes and hmnan epidermoid carcinoma cells, green tea induces apoptosis only in cells, but no keratinocytes. For endothelial cells, green tea inhibits endothelial cell proliferation cancer melanocytes and fibroblast, it is little known. We investigated the effects of green tea polyphenol(GTP) and the major constituents, (-)-epieallocatechin-3-gallate(EGCG) on the proliferation in the cultured human keratinocytes, fibroblasts, melanocytes, endothelial cells and human epidermoid carcinoma cells(A431 cells). The prolifertive response was studied by the cell count and the uptake of vitiated thymidine after 48 hours of treatment. And we also observed apoposs using TUNEL staining. The results were as follows; Number of living cells were significantly decreased(p<0,05) in cultured human epidermal cells, melanocytes, fibroblasts, endothelial cells and A431 cells treated with different concentration of GTP and EGCG for 48 hours. The cells were decreased dose dependently as increase of concentration of the extracts. The [3H]thymidine incorporation w significantly decreased(p<0.05) in cultured human epidermal cells, melanocytes, fibroblasts, endothelial cells and A431 cells treated with different concentration of GTP and EGCG for 48 hours. The dose dependent inhibition was also seen in [^3H]thymidine incorporation by green tea extracts. Cell proliferation was significantly more inhibited(p<0.05) by EGCG compared to GTP in the same concentration. We