全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

shRNA对大鼠心肌细胞牛磺酸转运体的抑制作用

DOI: 10.3969/j.issn.1006-267x.2013.11.017

Keywords: H9c2细胞,RNA干扰,牛磺酸转运体,细胞增殖

Full-Text   Cite this paper   Add to My Lib

Abstract:

本试验旨在研究短发夹RNA(shRNA)对大鼠心肌细胞牛磺酸转运体(TauT)的抑制作用。根据已克隆的大鼠心肌TauT基因序列设计并构建了3个shRNAs(shRNA1#、shRNA2#、shRNA3#)及1个阴性对照shRNA表达载体,利用脂质体LipofectamineTM2000将构建好的重组质粒shRNA1#、shRNA2#、shRNA3#及阴性对照质粒分别转染H9c2细胞,细胞分为对照组(未转染组)、shRNA-Neg组(阴性对照组)和3个转染组(shRNA1#组、shRNA2#组、shRNA3#组),每组3个重复。应用四唑盐(MTT)法检测H9c2细胞增殖情况,实时荧光定量PCR检测TauTmRNA表达水平。结果表明,与阴性对照组和未转染组比较,shRNA1#质粒仅在转染细胞后的24h极显著降低了TauTmRNA表达水平(P<0.01),shRNA2#质粒在转染细胞后的24、48、72h均极显著降低了TauTmRNA表达水平(P<0.01),shRNA3#质粒则是在转染细胞后的72h极显著降低了TauTmRNA表达水平(P<0.01);MTT检测结果表明,转染TauTshRNA1#、shRNA2#、shRNA3#入H9c2细胞24、48、72h后,细胞增殖均未受到显著影响(P>0.05)。上述结果提示,shRNA2#重组质粒能有效抑制TauT基因的表达,且短期内未影响H9c2细胞增殖,为研究TauT基因及揭示牛磺酸在心肌细胞代谢中的作用奠定基础。

References

[1]  WARSKULAT U,FL?GEL U,JACOBY C,et al.Taurine transporter knockout depletes muscle taurine levels and results in severe skeletal muscle impairment but leaves cardiac function uncompromised[J].The Journal of the Federation of American Societies for Experimental Biology,2004,18(3):577-579.
[2]  YAMORI Y,NARA Y,IKEDA K,et al.Is taurine a preventive nutritional factor of cardiovascular diseases or just a biological marker of nutrition?[J].Advances in Experimental Medicine and Biology,1996,403:623-629.
[3]  ITO T,OISHI S,TAKAI M,et al.Cardiac and skeletal muscle abnormality in taurine transporter-knockout mice[J].Journal of Biomedical Science,2010,17(Suppl.1):S20.
[4]  XU Y J,ARNEJA A S,TAPPIA P S,et al.The potential health benefits of taurine in cardiovascular disease[J].Experimental and Clinical Cardiology,2008,13(2):57-65.
[5]  HUXTABLE R J.Physiological actions of taurine[J].Physiological Reviews,1992,72(1):101-163.
[6]  SHI Y R,BU D F,QI Y F,et al.Dysfunction of myocardial taurine transport and effect of taurine supplement in rats with isoproterenol-induced myocardial injury[J].Acta Pharmacological Sinica,2002,23(10):910-918.
[7]  HELLER-STILB B,VAN ROEYEN C,RASCHER K,et al.Disruption of the taurine transporter gene (taut) leads to retinal degeneration in mice[J].The Journal of the Federation of American Societies for Experimental Biology,2002,16(2):231-233.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133