摘要
目的研究EMRE分子在高糖/高脂(high glucose and high fat,HGHF)培养的大鼠胚胎心肌细胞H9C2中的表达变化及其在HGHF诱导的心肌细胞凋亡中的作用。方法 H9C2细胞培养分为4组:即正常+si Ctrl组(培养基中含5 mmol/L葡萄糖、20 mmol/L甘露醇和si Ctrl,培养24 h)、正常+si-EMRE组(培养基中含5 mmol/L葡萄糖、20 mmol/L甘露醇和si-EMRE,培养24 h)、HGHF+si Ctrl组(培养基中含25 mmol/L葡萄糖、500μmol/L软脂酸钠和si Ctrl,培养24 h)及HGHF+si-EMRE组(培养基中含25 mmol/L葡萄糖、500μmol/L软脂酸钠和si-EMRE,培养24 h)。用qRT-PCR及Western blot检测EMRE在各组细胞中的表达;用流式细胞术(FCM)分析各组心肌细胞的凋亡;用Western blot检测caspase3与caspase9表达;用荧光探针DCFH-DA检测各组心肌细胞内ROS水平。结果HGHF诱导心肌细胞EMRE表达升高(mRNA及蛋白水平);EMRE促进了HGHF诱导的心肌细胞凋亡及ROS的产生;用si-RNA干涉EMRE表达后,由HGHF诱导的细胞凋亡及ROS产生明显减少。结论 EMRE在HGHF诱导的心肌细胞凋亡中发挥重要的促进作用。
AIM To investigate the expression of EMRE and its regulatory role in apoptosis of H9C2 cells induced by high glucose and saturated fatty acids （HGHF）. METHODS H9C2 cells were divided into four groups： control ＋ siCtrl group （5 mmol/L glucose, 20 mmol/L mannitol and siCtrl, incubated 24 h）, control ＋si-EMRE group （5 mmol/L glucose, 20 mmol/L mannitol and si-EMRE, incubated 24 h）, HGHF ＋siCtrl group （25 mmol/L glucose, 500 μmol/L palmitate sodium and siCtrl, incubated 24 h）, and HGHF ＋ si-EMRE group （25 mmol/L glucose, 500 μmol/L palmitate sodium and si-EMRE, incubated 24 h）. The mRNA and protein expression levels of EMRE were detected by qRT-PCR and Western blotting. Cell apoptosis was analyzed using flow cytometry, the expressions of caspase3 and caspase9 were detected by Western blotting and the intracellular ROS levels were assayed using fluorescence probe DCFH-DA. RESULTS HGHF induced up-regulation of EMRE in H9C2 cells both at mRNA and protein levels. EMRE was critical for HGHF-induced cell apoptosis and ROS production. Knocking down of EMRE using si-RNA significantly inhibited HGHF-induced cell apoptosis and ROS production. CONCLUSION EMRE is critical for HGHF-induced cell apoptosis.
出处
《心脏杂志》
CAS
2016年第3期285-288,共4页
Chinese Heart Journal
关键词
EMRE
高糖高脂
心肌细胞
活性氧簇
EMRE
high glucose and saturated fatty acids
H9C2
cardiomyocyte
ROS
