应激心肌细胞中c-Jun氨基末端激酶对TR3磷酸化水平的调控

目的观察应激心肌细胞中JNK的磷酸化水平及其对核转录因子TR3磷酸化水平的影响。方法采用应激心肌细胞模型,以10-5mol/L糖皮质激素(GC)干预心肌细胞;采用免疫印记法检测细胞中JNK和TR3蛋白的磷酸化水平;应用JNK磷酸化的化学抑制剂SP600125观察JNK磷酸化的抑制对TR3磷酸化水平的影响。用Image Master誖2D Elite(Version 3.10)软件分析蛋白条带的灰度值;用SPSS12.0软件包对灰度值进行统计学分析。结果选取对照组的p-JNK蛋白灰度值(0.99)相比,GC干预心肌细胞10min、20min、40min、60min、90min时的p-JNK蛋白灰度值升高(分别为1.26、1.31、1.29、1.35、1.36)。选取对照组的p-TR3蛋白灰度值(0.83)相比,GC干预心肌细胞40min、60min、90min时的p-TR3蛋白灰度值升高(1.09、1.16、1.02)。与GC干预组的p-JNK蛋白灰度值(1.55)相比,SP600125+GC组的p-JNK蛋白灰度值降低(1.21);与GC干预组的p-TR3蛋白灰度值(1.67)相比,SP600125+GC组的p-TR3蛋白灰度值降低(1.03)。结论在应激心肌细胞中,JNK活化,磷酸化水平升高,并进而促使核转录因子TR3发生磷酸化。

Regulation of JNK to TR3 Phosphorylation in Stressed Cardiomyocytes

Objective To study the level of JNK phosphorylation and its influence on TR3 phosphorylation, to illuminate molecular mechanism of its modulating TR3 in apoptosis occurring in stressed cardiomyocytes. Methods The cell model of tress- induced cardiomyocytes injury was established in vitro by using glucocorticoid of 10-s mol/L. The western blotting （WB） method was used to investigate the levels of JNK and TR3 proteins phosphorylation. Then using SP600125, the selective inhibitor of JNK phosphorylation, to observe the levels of TR3 phosphorylation. Optical density data of proteins was detected by using Image Master 2D Elite （Version 3.10） software and analyzed with SPSS 12.0 software. Results Compared with the control group （0.99）, the optical density data of p-JNK protein in glucocorticoid-treated group （GC group） （1.26,1.31,1.29,1.35,1.36） are greatly upregulated. Comparing with the control group（0.83）, the optical density datas of p-TR3 protein in 10-5 mol/L glucocorticoid disposed groups （1.09,1.16,1.02） are greatly advanced. Comparing with the glucocorticoid interfered wih group （1.55）,the optical density datas of p-JNK protein in the group pre-treated with SP600125 （1.21） is reduced distinctly. Comparing with the glucocorticoid interfered wih group （1.67）,the optical density dalas of p-TR3 protein in the group pre-treated with SP600125 （1.03） decrease apparently. Conclusion In stressed cardiomyocytes, JNK phosphorylate greatly and then phosphorylate TR3.