中国老年保健医学
中國老年保健醫學
중국노년보건의학
CHINESE JOURNAL OF GERIATRIC CARE
2013年
6期
25-26
,共2页
siRNA%大鼠%心肌细胞%沉默效果
siRNA%大鼠%心肌細胞%沉默效果
siRNA%대서%심기세포%침묵효과
siRNA%Rat%Cadiocyte%Silence effect
目的： Mg2＋是活细胞中含量最丰富的二价阳离子之一，是多种酶的辅助因子，还是维持大分子有活性的构象，脂类和磷酸肌醇类第二信使的调控，多种离子通道和转运体的调控所必需的在哺乳动物细胞生命活动中起着重要作用。临床观察表明，心肌细胞内游离Mg2＋浓度过低时会导致心律失常等现象。为了深入研究MagT1基因在心肌细胞中的功能，本研究试图确定沉默效果好的MagT1 siRNA，为后续实验奠定基础。方法体外分离大鼠心肌细胞，并检验心肌细胞的纯度；委托上海吉玛生物技术公司设计4对siRNA，与转染试剂混合并转染进大鼠心肌细胞，使用RT-PCR方法检测siRNA沉默MagT1基因的效果。结果经过RT-PCR的检测，发现4对siRNA中只有1对siRNA沉默效果最好，序列为5′-GAUUGGACUUGUUGUGUUATT-3′；5′-UAACACAACAAGUCCAAUCTT-3′。结论找到了沉默效果好的siRNA，为使用RNA干扰方法深入研究MagT1的功能奠定了基础。
目的： Mg2＋是活細胞中含量最豐富的二價暘離子之一，是多種酶的輔助因子，還是維持大分子有活性的構象，脂類和燐痠肌醇類第二信使的調控，多種離子通道和轉運體的調控所必需的在哺乳動物細胞生命活動中起著重要作用。臨床觀察錶明，心肌細胞內遊離Mg2＋濃度過低時會導緻心律失常等現象。為瞭深入研究MagT1基因在心肌細胞中的功能，本研究試圖確定沉默效果好的MagT1 siRNA，為後續實驗奠定基礎。方法體外分離大鼠心肌細胞，併檢驗心肌細胞的純度；委託上海吉瑪生物技術公司設計4對siRNA，與轉染試劑混閤併轉染進大鼠心肌細胞，使用RT-PCR方法檢測siRNA沉默MagT1基因的效果。結果經過RT-PCR的檢測，髮現4對siRNA中隻有1對siRNA沉默效果最好，序列為5′-GAUUGGACUUGUUGUGUUATT-3′；5′-UAACACAACAAGUCCAAUCTT-3′。結論找到瞭沉默效果好的siRNA，為使用RNA榦擾方法深入研究MagT1的功能奠定瞭基礎。
목적： Mg2＋시활세포중함량최봉부적이개양리자지일，시다충매적보조인자，환시유지대분자유활성적구상，지류화린산기순류제이신사적조공，다충리자통도화전운체적조공소필수적재포유동물세포생명활동중기착중요작용。림상관찰표명，심기세포내유리Mg2＋농도과저시회도치심률실상등현상。위료심입연구MagT1기인재심기세포중적공능，본연구시도학정침묵효과호적MagT1 siRNA，위후속실험전정기출。방법체외분리대서심기세포，병검험심기세포적순도；위탁상해길마생물기술공사설계4대siRNA，여전염시제혼합병전염진대서심기세포，사용RT-PCR방법검측siRNA침묵MagT1기인적효과。결과경과RT-PCR적검측，발현4대siRNA중지유1대siRNA침묵효과최호，서렬위5′-GAUUGGACUUGUUGUGUUATT-3′；5′-UAACACAACAAGUCCAAUCTT-3′。결론조도료침묵효과호적siRNA，위사용RNA간우방법심입연구MagT1적공능전정료기출。
Objective Mg2+is one of the most abundant cations in living cells which is cofactor of a variety of enzymes and maintained the conformation of the macromolecules activity ,regulated lipids and phosphoinositide second messenger.Mg2+plays im-portant role in mammal′s life activity which is necessary to the regulation of many kinds of ion channels and transporters .Clinical ob-servation showed that arrhythmia phenomenon appeared when the concentration of Mg 2+which in myocardial cells is low.In order to study the function of MagT 1 gene in myocardial cells deeply ,this study tried to determined the MagT 1 siRNA which with good silence effect,and to lay the foundation for follow -up study.Methods Isolated rat myocardial cells in vitro and checked the purity of myo-cardial cells ,entrusted Shanghai Jima Biotech Corp to designed four pairs of siRNA and mixed then dyed into rat cadiocyte ,tested the effect of siRNA silenced MagT 1 gene by the method of RT -PCR.Results Only one pairs of siRNA silence effect were found best though the test of RT -PCR,the sequence were 5′-GAUUGGACUUGUUGUGUUATT-3′;5′-UAACACAACAAGUCCAAUCTT-3′.Conclusion The siRNA that have better silence effect was found which laid the foundation for study the function of MagT 1 deep-ly by the method of RNA interference.
