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        • 다형홍반증에서 각질형성세포의 ICAM-1, TNF-α 및 IL-1α 발현에 관한 연구

          황규왕,정현,이종석,박준홍,부태성,이성열 순천향의학연구소 1998 Journal of Soonchunhyang Medical Science Vol.4 No.1

          The pathogenetic mechanisms involved in the developement of erythema multiforme(EM) are still largely unknown. The histologic and immunopathologic changes in erythema appear to be due in part to cellular immune mechanisms with the cytotoxic T cell. Especially cytotoxic T cell/ICAM-1 positive-keratinocyte adherence plays an important role in the pathogenesis of EM. In this study, we investigated the expression of TNF-a, IL-1 a by immunohistochemical stain and ELISA. In skin tissue of EM, strong basal cell expression of ICAM-1, TNF-a and IL-1 a was found, and we detected more TNF-a from supernatants of cultured human keratinocytes with serum of EM patient than normal control. Our results may suggest possible relationship between the pathogenesis of EM and cytokine from keratinocyte.

        • 인체 흑색종 세포군에서의 Vascular Endothelial Growth Factor(VEGF) 분비

          김현조,황규왕,조문균,박영립,이종성,이성열 순천향의학연구소 2001 Journal of Soonchunhyang Medical Science Vol.7 No.2

          Backgroung : Vascular endothelial growth factor(VEGF) is an angiogenic cytokine expressed by many human and animal tumors. Ultraviolet radiation(UVR) has been causally linked to melanoma initiation and progression. UVR is a known melanocyte mutagen and induces specific mutations in cellular oncogenes such as p53 and ras. Activated H-ras activates tumor angiogenesis in other cell types, and angiogenic switching is accompanied by up-regulation of VEGF. Objective : The purpose of this study was to determine whether VEGF release from human melanoma cell lines was altered by activating ras mutations or by inactivation of p53 induction and whether VEGF release of these transfected melanoma cell lines was changed after UVR radiation. Results : 1. Release of VEGF from menolayer culture of melanoma cells was significantly higher in cells with activating ras mutations and lower in cells with expression of dominant negative p53. 2. In spheroid culture, dominant negative p53 decreases VEGF release in WM35 cells, but activating ras did not affect VEGF release significantly. 3. The control cell line(WM35Neo) had higher VEGF release in the spheroid culture than in the monolayer culture, and dominant negative p53 or activating ras did not affect VEGF release significantly between these two culture systems. 4. Higher cell density increases VEGF release in all tested melanoma cell lines was shown in both spheroid and monolayer culture. This cell density effect was strongest in cells with activated ras in monolayer culture. 5. UVR inhibits VEGF release in all tested melanoma cell lines, and this effect was stronger in monolayer culture than in spheroid culture. The effect of UVR on VEGF release was significantly lower in the DNp53 transfectants. 6. UVR decreased the cell viability in a dose dependent manner regardless of the cell density or cell culture system. Conclusion : VEGF release in human melanoma was inhibited by ultraviolet radiation at cytotoxic doses, and this effect is stronger in monolayer culture than in spheroid culture. VEGF release from melanoma cell lines was also influenced by p53, ras and cell growth conditions in a complex manner.

        • 자외선 등의 자극에 의한 인체 멜라닌세포의 타이로신 프로테인 키나제 수용체 발현에 관한 연구

          이정열,황규왕,이종석,김영근 순천향의학연구소 1998 Journal of Soonchunhyang Medical Science Vol.4 No.1

          Human melanocytes express the p 75 nerve growth factor(NGF) receptors and tyrosine protein kinase receptors(trk) in vitro. Melanocytes are induced to express the p75 NGF receptor in vitro by a variety of phamacologic and physiologic stimuli including UV light. Melanocytes dendricity and migration are regulated in part by NGF. Cultured human epidermal keratino- cytes produce NGF. The purpose of the study was to investigate the influence of UVB and TPA(phorbol 12-tetradecanoate 13-acetate) stimuli in pure melanocytes group and the influence of UVB stimuli in keratinocytes/melanocytes coculture group in the expression of trk A, respectively. Normal human keratinocytes and melanocytes were established from neonatal foreskin. Keratinocytes and melanocytes were cultured by modified Medium 154 and F-10, respectively. Pure melanocytes group and keratinocytes/melanocytes coculture group were respectively exposed to UVB 50 mJ/㎠. pure melanocytes group were treated for 1 day with TPA 50, 100 ng/ml. the expression of trk A in melanocytes with 50 ng/ml TPA was induced about 3 times higher than unstimulated control. But. TPA induction of trk A in melanocytes is not dose-dependent. The expression of trk A in pure melanocytes group and melanocytes/keratinocytes cocultures group with UVB 50 mJ/㎠ was induced about 4-5 times than unstimulated control. We conclude that stimulation with TPA and UVB induces trk A in melanocytes and UVB effect is further potentiated by the presence of keratinocytes in culture.

        • 우리나라 남해안과 제주 근해에서 자생하는 해조류 추출물의 Melanogenesis에 대한 억제 효과

          송무현,황규왕 순천향의학연구소 2001 Journal of Soonchunhyang Medical Science Vol.7 No.2

          Backgroung: Many people have a great interest on skin pigmentation due to melnin production which is caused by UV exposure, hormone, physical and chemical injury from the cosmetic viewpoint. Therefore, need of new depigmenting agent which has safety and potency has been recently increasing. Traditionally, sea weeds have been considered as whitening agents in Korea. We screened sea weeds which was grown in southern parts of Korean sea whether they had tyrosinase inhibition activities by mushroom tyrosinase inhibition assay. Objective:Our purpose was to examine the inhibitory effects of Seaweed extracts on tyrosinase activity and melanin synthesis and compating with other depigmenting agents such as kojic acis, arbutin, licorice extract. Methods: Tyrosinase activity was determined by spectrophotometry, the effects of whitening agents(Kojic acid, arbutin, licorice extracts, Seaweed extracts) on mushroom tyrosinase was compared by measuring the IC_(50), the concentration of the compound at which half of the original tyrosinase activity was inhibited. B-16 melanoma cells were cultured in a pair of 6-well plate 37℃, 5% CO_(2), with DMEM plus 10% FCS contaning of various concentrations of Seaweed extracts. After 24 hours B-16 cells were harvested with trypsinization. B-16 cell pellets were used for measurement of mRNA expression for tyrosinase, TRP-1 and TRP-2 by RT-PCR. Results: 1. Seaweed extracts has relatively strong inhibitory effect on tyrosinase according to increment of concentration. 2. Seaweed extracts has relatively strong inhibitory effect on tyrosinase as compared with kojic acid, arbutin, licorice extract. 3. Seaweed extracts showed inhibitory effect on Tyrosinase and TRP-2 gene transcription but didn't showed inhibitory effect on TRP-1 gene transcription. Conclusion This study provided that Seaweed extracts has strong inhibitory effects aganise melanogenesis. The results suggest that Seaweed extracts will be able to use as a new whitening agent.

        • 정상 인체 멜라닌세포에서 endothelin-1 receptor antagonist가 melanin생성에 미치는 영향에 관한 연구

          박진우,이종석,황규왕,김정하 순천향의학연구소;Soonchunhyang Medical Research Institute 2000 Journal of Soonchunhyang Medical Science Vol.6 No.1

          In the normal skin, melanocytes maintain their constant population and their melanogenic activity in steady state, but respond to several stimulus such as UV radiation and chemical irritation, to accentuate proliferation and differentiation, leading to hyperpigmentation of the skin. Human Keratinocytes produce and secrete endothelin-1, which can be strong mitogens for human melanocytes. The paracrine linkage of endothelin-1 between keratinocytes and melanocytes suggested that endothelin-1 is intrinsic mediators for human melanocytes in UVB-induced pigmentation. The purpose of this study is to investigate the new whitening agent, which improve the hyperpigmentation by inhibiting the ET-1. So, we add ET-1 and five kinds of ETRA into the cultured normal human melanocyte and then three days later, each ETRA on ET-1 was measured melanocyte number and inhibitory effect of melanin synthesis, respectively. In this study, ETRA has a inhibitory effect on ET-1 induced melanocyte proliferation and melanogenesis. ETRA I has a strong inhibitory effect on melanocyte proliferation and ETRA Ⅱ has a strong inhibitory effect melanogenesis. western blotting results shows negative. It may be due to the processing error of protein denaturing or low antigen -antibody affinity. But, we also confirm that ETRA Ⅱ has a inhibitory effect of tyrosinae synthesis by RT-PCR

        • 무색소성 색소 실조증(Ito 저색소증) 1예

          방성원,이정열,황규왕,정현,김영근 순천향의학연구소 1995 Journal of Soonchunhyang Medical Science Vol.1 No.1

          Incontinentia pigmenti achromians(IPA) or Hypomelanosis of Ito is a very rare neurocutaneous syndrome which is often associated with cutaneous, cerebral, musculoskeletal and ocular abnormalities. It is assumed that it is autosomal dominant of inheritance in character. There is also a relationship between IPA and incontinentia pigmenti(IP). However, the etiology of IPA is not known. We describe a 4-year-old girl with myopia, esotropia and retinal pigmentary degeneration in addition to the hypopigmented skin lesion of IPA, and reviewed clinically the cases of IPA in this article.

        • 정상 인체 멜라닌세포 및 B-16 흑색종 세포주에서 상지(桑枝) 추출물의 티로시나제 활성 억제 효과에 관한 연구

          송무현,황재영,박영립,황규왕 순천향의학연구소 2000 Journal of Soonchunhyang Medical Science Vol.6 No.2

          Background : Recently, the undesirable effects of UV exposure have been increasing because of destruction of ozone layer and excessive solar exposure in enjoying leisure. Therefore, the chance to have uneven skin pigmentations has been increasing. To keep away from the unwanted skin pigmentation, melanogenesis inhibitors have been developed for use in cosmetic preparations for the purpose of skin whitening. Plant extracts having an inhibitory effect in melanin synthesis may be a good choice as a cosmetic ingredient because they have relatively fewer side effects. Objective : In this study, the inhibitory effects of ramulus mori (young twigs of Morus alba L) on tyrosinase activity were investigated cultured normal human melanocytes and B-16 melanoma cells by using enzyme assay and RT-PCR. Methods : Tyrosinase activity was determined by spectrophotometry. The effects of whitening agents (kojic acid, arbutin, licorice extracts and ramulus mori extracts) on mushroom tyrosinase was compared by measuring the IC_(50), the concentration of the compound at which half of the original tyrosinase activity is inhibited. Normal human melanocytes taken from neonatal foreskin and B-16 melanoma cells, tyrosinase activity inhibition was measured by spectrophotometry. We observed tyrosinase volume in B-16 melanoma cells by using PT-PCR. Results : Ramulus mori extracts showed strong inhibitory effect on tyrosinase activity in both normal human melanocytes and B-16 melanoma cells. We also confirm that they have a inhibitory effect on tyrosinase expression in the RT-PCR. Conclusion : This study showed that ramulus mori extracts had strong inhibitory effect against tyrosinase activity. The results suggests that ramulus mori extracts can be used as a new whitening agent.

        • 아토피 피부염 환자에서 혈청내 Cu와 Zn치 및 Cu/Zn비에 관한 연구

          이성열,정현,이종석,황규왕 순천향의학연구소 1998 Journal of Soonchunhyang Medical Science Vol.4 No.1

          아연과 구리는 피부를 포함하는 인체의 거의 모든 기관에서 일어나는 대사과정에 작용하는 metallonenzyme의 보조인자로 상당히 중요한 역할을 수행한다. 저자들은 정상대조군 10명, 전신성 아토피성 피부염 환자 16명을 대상으로 atomic absorption spectrometry를 사용하여 혈청 아연과 구리치를 측정하였고 그결과로 혈청 구리/아연 비를 구하였다. 그 결과 혈청 구리치는 대조군에 비하여 아토피성피부염 환자에서 통계적으로 의의있게 높게 나타났다. 이는 아마도 아토피성 피부염에 의한 염증반응으로 이차적으로 ceruloplasmin이 증가되어 나타난 것으로 생각되나 이 것이 명확히 규명된 것은 아니다. 혈청 아연치는 약간 감소하였으나 통계적인 유의성은 없었고 혈청 구리/아연 비의 값은 약간 증가되었으나 역시 통계적 유의성을 찾을 수는 없었다.

        • 해조류에서 추출한 SCH-T3의 elastase 억제

          오경석,박영립,이종석,황규왕,염윤기 순천향의학연구소 2001 Journal of Soonchunhyang Medical Science Vol.7 No.1

          Background & Objective: Upon UV irradiation of skin, matrix metalloproteinases are induced and then degrade collagen and elastin which provide the structural integrity of skin. The purpose of this study is to investigate the inhibition of elastase by SCH-T3 extracted from marine plants. Method: From gathered seaweeds, which are Laminariacea and Alariaceae in Phaeophyta, In Korea water, 21 materials were extracted. To determine the optimal condition for the extraction of SCH-T3, the yields of SCH-T3 was measured compared at various solvents, concentrations, and extraction time. Finally, the inhibitory effect of SCH-T3 on elastase was measured under various conentrations of SCH-T3. Results: Among 21 extracts, SCH-T3 had excellent elastase inhibitory effect. The yields of SCH-T3 was highest with the use of 60% and 70% ethanol as sovents, at 12 hours extraction, and with increasing temperature. Conclusion: These results suggest that SCH-T3, extracted from marine plants, has an inhibitory effect on elastase in vitro and may be used in prevention and treatment of photoaging in the future.

        • 자외선 조사에 의한 인체 각질형성세포 세포고사 방어인자에 관한 연구

          박수홍,박준홍,이종석,황규왕,김계정 순천향의학연구소 1998 Journal of Soonchunhyang Medical Science Vol.4 No.2

          Back ground: Human skin is continuously exposed to UV irradiation. Ultraviolet irradiation of human skin cause sunburn cell which is relevant to the apoptosis of keratinocytes. In the epidermis, apoptosis inducing factors and anti-apoptotic factors probably exist to maintain the integrity of keratinocytes. Objective: The purpose of this study was to investigate the exsistence of apoptosis inducing factors and anti-apoptotic defence factors by evaluation of UV induced apoptosis in cultured human keratinocyte and other keratinocyte cell lines(A-431 cells, KB cells) and its susceptibility of UV induced apoptosis in various conditions. Method: In this study, the percentages of apoptosis, necrosis and cell viability of irradiated human keratinocytes and othe keratinocyte cell lines by MMT assay and AO/EO stain. The percentages of those were measured before UVB irradiation and 8, 24, 48 hours after UVB irradiation. Also, the same evaluations were performed with irradiated human keratinocytes cultured without growth factor and with enough growth factors, both results were compared with each other. And the effect of cycloheximide, a protein synthesis inhibitor was evaluated. Also that of aurintricarboxylic acid(ATA), an inhibitor of endonucleases which play an important role in inducing apoptosis of human keratinocytes was evaluated. Result: Human keratinocytes and other keratinocyte cell lines(A-431 cells, KB cells) were cultured in vitro developed maximal apoptosis 48 hours after irradiation, keratinocytes were more resistant to UV induced apoptosis than the others. The withdrawal of growth factors from keratinocyte and addition of cycloheximide decreased the cell survival rate following UV irradiation and increased the induction of apoptosis. And ATA inhibited UV induced apoptosis. Conclusion: These results indicate that human keratinocytes have both anti-apoptotic factors and apoptosis inducing factors to maintain the homeostasis. And survival signals mediated through growth factors or cellular proteins are responsible for the resistance to apoptosis observed in keratinocytes in vitro.

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