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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2010, Vol. 30 Issue (09): 1-6    DOI: Q2
研究报告     
TNF-a在乳鼠心肌细胞中诱导HSP70蛋白表达的研究
黄银霞,魏英杰**,崔传珏,张晓玲,李君
中国医学科学院阜外心血管病医院 卫生部心血管疾病再生医学重点实验室 北京 100037
Tumor necrosis factor-a-induced expression of heat shock protein 70 in Neonatal rat ventricular cardiomyocytes
HUANG Yin-xia,WEI Ying-jie,CUI Chuan-jue,ZHANG Xiao-ling,LI Jun
Key Laboratory of Cardiovascular Regenerative Medicine, Ministry of Health, Cardiovascular Institute and Fu-Wai Hospital, PUMC and CAMS, Beijing 100037, China  
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摘要:

热休克蛋白70 (HSP70) 在细胞修复、存活和维持细胞正常功能方面有着重要作用。作为分子伴侣,它起着心肌保护的作用。已经对重症心脏病人的心肌组织进行了蛋白组学研究,得到了HSP70在心衰病人心肌组织中较正常人心肌组织表达升高的结论,并且在血液中得到了进一步的验证。在进一步的离体细胞实验中用不同剂量的肿瘤坏死因子-alpha (TNF-α) 刺激乳鼠心肌细胞,以观察不同时间点HSP70的动态表达情况。培养乳鼠心肌细胞,分别对细胞进行热休克(42 ℃)、TNF-α和缺血缺氧处理,在不同的时间点收获细胞,以观察HSP70的动态表达情况。用免疫化学、ELISA以及Western blotting的方法对HSP70蛋白进行分析。结果表明,在正常对照细胞中基本没有阳性信号出现,而在经缺血缺氧、热休克(42 ℃)以及TNF-α处理的细胞中有明显的阳性表达。以上研究首次在乳鼠心肌细胞中证明TNF-α诱导的HSP70表达具有时间和浓度依赖性。通过运用TNF-α对HSP70蛋白表达影响的研究,初步推断HSP70的表达模式,为体内诱导产生HSP70从而发挥心肌保护作用的研究提供一定的理论基础。

关键词: HSP70缺血缺氧肿瘤坏死因子-alpha新生乳鼠心肌细胞    
Abstract:

Heat shock protein 70 is essential for cellular recovery, survival and maintenance of cellular function. Heat shock protein 70 function as molecular chaperons in protein folding and it is a role of cardioprotection against various stresses. We have studied the different proteins expression of tissues from failing hearts using proteomics before. We found that HSP70 was highly expressed in the tissues and blood samples from heart failure patients compaired with non-failing hearts. For exploiting the use of HSP70 as a therapeutic marker, we sthudy the HSP70 expression using Neonatal rat ventricular cardiomyocytes(NRVM) by teating with TNF-a with various concentrations and time points in vitro. Primary cultures were prepared from neonatal rat ventricular myocytes. Cultured cardiomyocytes were incubated with TNF-a at doses of 40, 200 and 800U /ml for a dynamic time point as 1, 6, 12 and 24 hours. NRVM treated with heat shock(42℃) were used as positive control. The expression of myocyte HSP70 was quantified by Western blot analysis and immunohistochemistry using a specific monoclonal antibody. Immunohistochemical analysis revealed HSP70 were nearly not visible in control cells. However, at 12 h post-hypoxia/SD, HSP70-immunoreactivity (IR) was evident in the cells. These studies constitute the initial demonstration that TNF-alpha exerts concentration- and time-dependent effects on the expression of HSP 70 in NRVM. The expression of HSP70 influenced by TNF-a can indicate the possibility of a pharmacological approach to HSP70 induction and cardiac protection, which may ultimately be of clinical relevance.

Key words: Heat shock protein 70    Hypoxia/Serum Deprivation    TNF-α    Neonatal rat ventricular cardiomyocytes(NRVM)
收稿日期: 2009-12-21 出版日期: 2010-08-25
基金资助:

国家“863”计划(2006AA02Z4B8)资助项目

通讯作者: 魏英杰     E-mail: weiyingjiefw@126.com
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引用本文:

黄银霞 魏英杰 崔传珏 张晓玲 李君. TNF-a在乳鼠心肌细胞中诱导HSP70蛋白表达的研究[J]. 中国生物工程杂志, 2010, 30(09): 1-6.

HUANG Yin-Xia, WEI Yang-Jie, CUI Chuan-Jue, ZHANG Xiao-Ling, LI Jun. Tumor necrosis factor-a-induced expression of heat shock protein 70 in Neonatal rat ventricular cardiomyocytes. China Biotechnology, 2010, 30(09): 1-6.

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https://manu60.magtech.com.cn/biotech/CN/Q2        https://manu60.magtech.com.cn/biotech/CN/Y2010/V30/I09/1

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