癌变·畸变·突变
癌變·畸變·突變
암변·기변·돌변
CARCINOGENSES,TERATOGENSIS AND MUTAGENESIS
2014年
6期
423-427,433
,共6页
崔宝弟%武扬%孙震晓
崔寶弟%武颺%孫震曉
최보제%무양%손진효
去甲斑蝥素%心肌细胞%缺血再灌注损伤%程序性细胞死亡因子4%microRNA-21
去甲斑蝥素%心肌細胞%缺血再灌註損傷%程序性細胞死亡因子4%microRNA-21
거갑반모소%심기세포%결혈재관주손상%정서성세포사망인자4%microRNA-21
norcantharidin%rat cardiac myocyte%myocardial ischemia-reperfusion injury%programmed cell death 4%microRNA-21
目的：探讨去甲斑蝥素(NCTD)抗大鼠心肌缺血再灌注损伤(IRI)的作用及其机制。方法：以大鼠乳鼠原代心肌细胞和大鼠心肌细胞系H9c2为研究对象，采用MTT法检测不同浓度(0.01、0.05、0.1、0.5、1和5μg/mL)NCTD预处理对IRI的心肌细胞活力的影响；流式细胞术检测NCTD预处理对IRI的心肌细胞凋亡的影响；Giemsa染色及台盼蓝染色观察NCTD预处理对IRI的心肌细胞形态的影响；Western blot检测NCTD预处理对IRI心肌细胞中程序性细胞死亡因子4(Pdcd4)表达情况；RT-qPCR检测NCTD预处理对IRI心肌细胞microRNA-21(miR-21)的表达情况。结果：NCTD对原代心肌细胞和H9c2心肌细胞IRI具有抑制作用(P<0.01)；通过NCTD预处理可以抑制IRI造成的细胞死亡并抑制IRI导致的Pdcd4蛋白表达上调及miR-21下调。结论：NCTD在0.1~0.5μg/mL浓度下可以抑制心肌细胞IRI，miR-21-Pdcd4信号通路参与心肌细胞IRI及NCTD抑制IRI的作用。
目的：探討去甲斑蝥素(NCTD)抗大鼠心肌缺血再灌註損傷(IRI)的作用及其機製。方法：以大鼠乳鼠原代心肌細胞和大鼠心肌細胞繫H9c2為研究對象，採用MTT法檢測不同濃度(0.01、0.05、0.1、0.5、1和5μg/mL)NCTD預處理對IRI的心肌細胞活力的影響；流式細胞術檢測NCTD預處理對IRI的心肌細胞凋亡的影響；Giemsa染色及檯盼藍染色觀察NCTD預處理對IRI的心肌細胞形態的影響；Western blot檢測NCTD預處理對IRI心肌細胞中程序性細胞死亡因子4(Pdcd4)錶達情況；RT-qPCR檢測NCTD預處理對IRI心肌細胞microRNA-21(miR-21)的錶達情況。結果：NCTD對原代心肌細胞和H9c2心肌細胞IRI具有抑製作用(P<0.01)；通過NCTD預處理可以抑製IRI造成的細胞死亡併抑製IRI導緻的Pdcd4蛋白錶達上調及miR-21下調。結論：NCTD在0.1~0.5μg/mL濃度下可以抑製心肌細胞IRI，miR-21-Pdcd4信號通路參與心肌細胞IRI及NCTD抑製IRI的作用。
목적：탐토거갑반모소(NCTD)항대서심기결혈재관주손상(IRI)적작용급기궤제。방법：이대서유서원대심기세포화대서심기세포계H9c2위연구대상，채용MTT법검측불동농도(0.01、0.05、0.1、0.5、1화5μg/mL)NCTD예처리대IRI적심기세포활력적영향；류식세포술검측NCTD예처리대IRI적심기세포조망적영향；Giemsa염색급태반람염색관찰NCTD예처리대IRI적심기세포형태적영향；Western blot검측NCTD예처리대IRI심기세포중정서성세포사망인자4(Pdcd4)표체정황；RT-qPCR검측NCTD예처리대IRI심기세포microRNA-21(miR-21)적표체정황。결과：NCTD대원대심기세포화H9c2심기세포IRI구유억제작용(P<0.01)；통과NCTD예처리가이억제IRI조성적세포사망병억제IRI도치적Pdcd4단백표체상조급miR-21하조。결론：NCTD재0.1~0.5μg/mL농도하가이억제심기세포IRI，miR-21-Pdcd4신호통로삼여심기세포IRI급NCTD억제IRI적작용。
OBJECTIVE: To explore the effects of norcantharidin (NCTD) on myocardial ischemia-reperfusion injury (IRI) and its possible mechanisms in rats. METHODS:The effect of different concentrations of NCTD on primary rat cardiomyocytes and H9c2 cells was examined by MTT assay. The apoptosis effect of NCTD pretreatment on IRI is detected by flow cytometry. Ischemia-reperfusion myocardial cell morphology of NCTD pretreatment was examined by Giemsa and Trypan blue staining. The expression of programmed cell death factor 4 (Pdcd4) in NCTD-pretreated ischemia-reperfusion myocardial cell was examined by Western blot. The expression of microRNA-21(miR-21) in NCTD-pretreated ischemia-reperfusion myocardial cell was assessed by RT-qPCR. RESULTS:The IRI of primary rat cardiomyocytes and H9c2 was inhibited by NCTD (P<0.01). Cell death caused by IRI was significantly inhibited by NCTD. NCTD could effectively inhibit Pdcd4 protein up-regulation and miR-21 down-regulation caused by IRI in rat cardiomyocytes. CONCLUSION:Myocardial IRI could be inhibited by NCTD treatment in concentration of 0.1-0.5μg/mL,and miR-21-Pdcd4 signaling pathways may participate in myocardial IRI and the inhibition of IRI by NCTD.
