去甲肾上腺素诱导大鼠心肌细胞NF-κB活化的研究

Effect of norepinepherine on the activity of NF-κB in cultured rat cardiomyocytes

目的观察去甲肾上腺素(NE)诱导大鼠心肌细胞NF-κB活化的影响,并探讨其作用机制。方法出生1-3d的SD大鼠,分离、培养心肌细胞,将培养4-5d呈亚融合状态下细胞随机分为6组(每组n=4):对照组、10-6mol/LNE组(NE1组)、10-7mol/LNE组(NE2组)、10-8mol/LNE组(NE3组)、10-5mol/L酚妥拉明+10-8mol/LNE组(Phe+NE组);10-5mol/L艾司洛尔+10-8mol/LNE组(Es+NE组)。NE1组、NE2组、NE3组细胞培养基内分别加入相应浓度的NE,Phe+NE组、Es+NE组在加入NE前30min分别加入10-5mol/L酚妥拉明、10-5mol/L艾司洛尔。加入NE后24h,采用流式细胞术(FCM)测定心肌细胞胞质内NF-κB表达百分比以反映其活化程度。结果与对照组相比,NE1组、NE2组、NE3组NF-κB活化程度升高,其中NE3组NF-κB活化程度最高(P<0.05);与NE3组比较,Phe+NE组、Es+NE组NF-κB活化程度降低(P<0.05)。结论α受体拮抗剂酚妥拉明和β1受体拮抗剂艾司洛尔均可单独完全阻断NE诱导的心肌细胞NF-κB活化作用,提示NE既可通过α受体又可通过β1受体来诱导心肌细胞NF-κB活化。

Objective To investigate the effect of norepinepherine（NE）on the activity of NF-κ3 in cultured rat cardiomyocytes and its mechanism. Methods The cardiomyocytes from 1 - 3 d SD rats were isolated and cultured. Twenty-four wells of cardiomyocytes were randomly divided into 6 groups： control group, 10^-6 md/L NE group, 10^-7 mol/L NE group, 10^-8 md/L NE group, Phe ＋ NE group（ 10^-5 md/L phentdamine ＋ 10^-8 md/L NE）, Es＋ NE group（ 10^-5 md/L esmdd ＋ 10^-8 md/L NE）. At rain 30 before administration of NE, phentdamine and esmolol were added into the culture fluid in Phe ＋ NE group and Es ＋ NE group, respectively. After administration of NE for 24 h, NF-κB expression was detected for evaluating the activation by flow cytometry （FCM）. Results The activation of NF-κB increased significantly in NE groups as compared to that in Phe ＋ NE group, Es ＋ NE group and control group（P 〈 0.05）, highest in 10^-8mol/L NE group. Conclusion Pre-administration of phentolamine and esmold could completely block NE-induced activation of NF-κB in cardiomyocytesl which might imply that the effects of NE on cardiomyocytes may be mediated together or single by α-adrenergic and β1-adrenergic receptors.