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박건재,김연정,김정근,김상민,이상엽,배장환,황경국,김동운,조명찬 대한심장학회 2012 Korean Circulation Journal Vol.42 No.1
Background and Objectives: The redox system is an important anti-oxidative system composed of thioredoxin, thioredoxin reductase,and peroxiredoxin (PRx). The fine details of PRx expression and its protective effects in various cells in cardiovascular tissue under oxidative stress created by hydrogen peroxide have not been fully elucidated. Subjects and Methods: Oxidative stress was induced by adding hydrogen peroxide at 0.25 mM for 2 hours to rat neonatal cardiomyocytes (rCMCs), rat vascular smooth muscle cells (rVSMCs), and human umbilical vein endothelial cells (HUVECs). Apoptosis was quantified by flow cytometry and the expression patterns of the six PRx isoforms were evaluated by western blotting in the three cell lines after hydrogen peroxide stimulation. Apoptosis and the cell survival signal pathway were evaluated by PRx1 gene delivery using lentiviral vector in hydrogen peroxide stimulated rCMCs versus green fluorescence protein gene delivery. Results: Hydrogen peroxide induced 25% apoptosis in rCMCs. Furthermore, the PRx1 and 5 isoforms were found to be overexpressed in hydrogen peroxide treated rCMCs, and PRx1 overexpression by gene delivery was found to reduce hydrogen peroxide induced rCMCs apoptosis significantly. In addition, this effect was found to originate from cell survival pathway modification. Conclusion: Hydrogen peroxide induced significant oxidative stress in rCMCs, rVSMCs, and HUVECs, and PRx1 overexpression using a lentiviral vector system significantly reduced hydrogen peroxide induced rCMCs apoptosis by upregulation of cell survival signals and downregulation of apoptotic signals. These findings suggest that PRx1 could be used as a treatment strategy for myocardial salvage in conditions of oxidative stress.
박건재,김연정,최은주,박노관,김기현,김상민,이상엽,배장환,황경국,김동원,조명찬 대한심장학회 2010 Korean Circulation Journal Vol.40 No.12
Background and Objectives: The thioredoxin (TRx) system is a ubiquitous thiol oxidoreductase pathway that regulates cellular reduction/oxidation status. Although endothelial cell (EC) hypoxic damage is one of the important pathophysiologic mechanisms of ischemic heart disease, its relationship to the temporal expression pattern of the TRx system has not yet been elucidated well. The work presented here was performed to define the expression pattern of the TRx system and its correlation with cellular apoptosis in EC lines in hypoxic stress. These results should provide basic clues for applying aspects of the TRx system as a therapeutic molecule in cardiovascular diseases. Subjects and Methods: Hypoxia was induced with 1% O2, generated in a BBL GasPak Pouch (Becton Dickinson, Franklin Lakes, NJ, USA) in human endothelial progenitor cells (hEPC) and human umbilical vein endothelial cells (HUVEC). Apoptosis of these cells was confirmed by Annexin-V: Phycoerythrin flow cytometry. Expression patterns of TRx; TRx reductase; TRx interacting protein; and survival signals, such as Bcl-2 and Bax, in ECs under hypoxia were checked. Results: Apoptosis was evident after hypoxia in the two cell types. Higher TRx expression was observed at 12 hours after hypoxia in hEPCs and 12, 36, 72 hours of hypoxia in HUVECs. The expression patterns of the TRx system components showed correlation with EC apoptosis and cell survival markers. Conclusion: Hypoxia induced significant apoptosis and its related active changes of the TRx system were evident in human EC lines. If the cellular impact of TRx expression pattern in various cardiovascular tissues under hypoxia or oxidative stress was studied meticulously, the TRx system could be applied as a new therapeutic target in cardiovascular diseases, such as ischemic heart disease or atherosclerosis.
가임기 여성의 알코올 섭취에 의한 세로토닌 조절이상 및 태아 이상발달 연구
이대연,박건재,김지연,김규희,정은애,김원호 한국알코올과학회 2013 한국알코올과학회 학술대회 Vol.2013 No.04
본 연구에서는 가임기 여성의 알코올 섭취가 세로토닌 호르몬 조절 및 임신, 출 산에 미치는 영향을 알아보기 위하여, 사람 나이 20대 초반에 해당하는 젊은 쥐 와 30대 후반-40대 초반에 해당하는 나이든 쥐로 그룹을 나누고 각각의 그룹을 알코올 섭취군과 대조군으로 나누어 조사한 결과 다음과 같은 결과를 얻을 수 있었다. • 젊은 쥐에 비해 나이든 쥐의 경우 임신 및 출산 능력이 현격히 감소하였 고, 이는 알코올 섭취에 의해 더욱 더 심화되었다. 반면, 나이든 쥐의 경우 태아의 사산율 및 기형아 출산율이 현격히 증가하였다. • 알코올 섭취한 젊은 쥐의 경우 배아 발달이 정상군에 비해 조금 늦었고 출산율의 감소가 나타났다. 반면 나이든 쥐의 경우 젊은 쥐에 비해 배아와 태아의 발달에 심각한 문제가 야기되었다. • 임신연령 증가 및 알코올 섭취에 의해 프로락틴 및 세로토닌 호르몬 분비 패턴 변화가 나타났다. • 췌장에서의 프로락틴과 세로토닌(5-HT), 관련 수용체들의 유전자 발현에 있어서 임신연령 증가 및 알코올 섭취가 미치는 효과가 뚜렷하였다. 결론적으로 본 연구진은 가임기 여성의 알코올 섭취는 임신과 관련된 호르몬 조절의 이상을 유발하고 이에 따라 임신 및 출산율을 떨어뜨리는 것은 물론 태 아의 이상발달을 유발할 가능성이 있음을 실험동물에서 확인할 수 있었으며, 더 욱이 이러한 현상은 임신연령이 높아질수록 더 증가하는 것을 확인하였다 . 이는 현재 임신부 연령이 점차 증가함과 동시에 여성의 음주율이 같이 증가하고 있는 우리나라의 상황에서 시사하는 바가 크며, 이러한 결과를 참고하여 임상연구 및 역학조사를 진행하고 관련된 보건대책을 수립하는 것이 시급하다고 여겨진다.