去甲斑蝥素抑制大鼠心肌细胞缺血再灌注损伤的研究

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5 μg/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0.1~0.5 μg/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。

Protective effect of norcantharidin on ischemia-reperfusion injur y in rat cardiomyocytes

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.