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Roles of Src-family kinase isoforms, Lyn, Fyn, Fgr, and c-Src on degranulation in RBL-2H3 mast cells
이준호,문세환,고나영,김지완,김도균,김주동,허억,최완수,Lee, Jun-Ho,Mun, Se-Hwan,Ko, Na-Young,Kim, Jie-Wan,Kim, Do-Kyun,Kim, Joo-Dong,Her, Erk,Choi, Wahn-Soo Korean Society of Life Science 2007 생명과학회지 Vol.17 No.3
흰쥐유래의 비만세포인 RBL-2H3 세포는 다양한 Src-family kinase를 발현한다. 현재까지의 연구결과에 의하면 비만세포의 초기 활성화에 Lyn kinase가 중요한 역할을 한다고 알려져 왔다. 그러나 그 세포에서 발현되는 다양한 다른 Src-family kinase의 역할은 불분명하다. 본 연구에서는 비만세포에서 다양한 Src-family kinase가 세포내 다른 곳에서 다양하게 발현되고 있다는 사실을 RT-PCR, immunoblotting 그리고 confocal microscopy 기법을 이용하여 증명하였다. 그 결과 Lyn 및 Fgr kinase는 세포막에 위치하고 c-Src 및 Yes kinase는 세포 내 과립에 존재하는 것을 알 수 있었다. 모든 Src-family kinase를 클로닝하고 과발현하여 탈과립 대한 영향을 평가하였다. 그 결과 fyn과 Fgr kinase는 비만세포에서 항원 유도의 탈과립을 증가시켰으며 반면 Lyn kinsae는 탈과립을 억제시키는 것을 확인할 수 있었다. 이러한 결과는 비만세포 초기 신호전달계에서 Fgr가 중요한 역할을 할 가능성을 제시한다. The rat RBL-2H3 mast cells contain various Src-family kinases. Previous reports with this cell line indicated that Lyn activation is an important initial signaling for the activation of the cells. However, the role and location of other Src-family kinase isoforms which are expressed in the cells are not clear. In this study, we now show that isoforms of Src-family kinases, Lyn, fyn, Fgr, c-Src, and Yes are differentially expressed and located differently in the cells as indicated by RT-PCR, immunoblotting analysis, and confocal microscopy. Lyn and Fgr were located on plasma membrane but on the other hand c-Src and Yes were located on intracellular organelle. All of Src-family kinases were cloned and overexpressed for investigating the roles of the isoforms. Overexpression of Fyn and Fgr, not Lyn and c-Src, stimulated Ag-induced degranulation in the cells. Our findings strongly suggest for the first time that each of Src-family kinase isoform can regulate differentially $Fc{\varepsilon}RI$-mediated signaling in RBL-2H3 mast cells.
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Macrophages로부터 IL-1β 분비 및 전사에 있어서 한국산 겨우살이 추출물 M11C (non-lectin components)의 효과
장성호,전명하,강태봉,문세환,이준호,성낙술,이성태,김종배,허억,Chang, Sung Ho,Jun, Myung Ha,Kang, Tae Bong,Mun, Se Hwan,Lee, Jun Ho,Seong, Nak Sul,Lee, Sung Tae,Kim, Jong Bae,Her, Erk 대한면역학회 2001 Immune Network Vol.1 No.2
Background: Korean mistletoe (Viscum album) extract has been found to posses immunostimulatory activity. In this study, Korean mistletoe extract, M11C (non-lectin components), was used to know whether this extract might activate mouse peritoneal macrophages to produce interleukin $1{\beta}$ (IL-$1{\beta}$). Methods: Hemagglutination assay was carried out to examine whether M11C contained a lectin or not. To know the effect of M11C on the production of IL-$1{\beta}$, the macrophages were treated by the M11C, and then collected the supernatant (M11C stimulated macrophages-conditioned media; MMCM). MMCM was analyzed for the IL-$1{\beta}$ quantification and mRNA expression by means of ELISA and RT-PCR, respectively. Results: Maximum effective dose and time of M11C on IL-$1{\beta}$ production from macrophages were $20{\mu}g/m{\ell}$ and 8 hours, respectively. This ELISA data was reconfirmed by immunoblotting assay. indicating that M11C is a good candidate for an immunomodulator. The dose and time dependent effects of M11C on the expression of IL-$1{\beta}$ mRNA from macrophages was also shown in expression of mRNA detected by RT-PCR. Treatment dose and time for the maximum expression of IL-$1{\beta}$ mRNA were $20{\mu}g/m{\ell}$ and 4 hours, respectively. Maximum gene expression of IL-$1{\beta}$ was much earlier than maximum production of it. Conclusion: As results, Korean mistletoe extract, M11C, may be used for an immunomodulator. This will be able to make up for and solve the problems caused by existent immunoagent with many adverse effects through many other studies in future including one molecule extraction.
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골관절염 연골세포에서 진행성당화종말생성물에 의한 기질단백분해효소 발현의 증가
나성수 ( Seong Su Nah ),최인영 ( In Young Choi ),문세환 ( Se Hwan Mun ),김용길 ( Yong Gil Kim ),문희범 ( Hee Bom Moon ),유빈 ( Bin Yoo ),이창근 ( Chang Keun Lee ) 대한류마티스학회 2007 대한류마티스학회지 Vol.14 No.1
Objective: Although increased expression of receptor for advanced glycation end products (AGE) in osteoarthritis (OA) has been reported, little is known concerning the role of AGEs in the pathogenesis of OA. This study was undertaken to determine the effect of AGEs on the regulation of matrix metalloproteinase (MMP) expressions and activities in human OA chondrocytes Methods: OA chondrocytes were treated with increasing doses of AGE-bovine serum albumin (AGE-BSA). The expressions of MMPs were determined by both enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis. The activities of MMPs were evaluated by both gelatin and casein zymography assays. In addition, electrophoretic mobility shift assay (EMSA) was employed to investigate the DNA binding activity of nuclear factor-kappa B (NF-κB) by AGE-BSA treatment. Results: The productions of MMP-1, -3, and -13 were significantly elevated by AGE-BSA in a dose dependent manner. The elevated activities of MMP-1, -3, and -13, and TNF-α by AGE-BSA were also observed. DNA binding activity of NF-κB was markedly increased by AGE-BSA treatment implicating possible involvement of NF-κB mediated pathway in the AGE-BSA induced MMP-1, -3, and -13, and TNF-α productions in OA chondrocytes. Taken together, this study demonstrates the stimulatory effect of AGE-BSA on the productions of MMPs and TNF-α and suggests the possible involvement of NF-κB mediated pathway in OA chondrocytes. Conclusion: These results suggest that AGE may play a role in pathogenesis of OA.
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