重组腺病毒转染心衰大鼠心肌细胞cAMP水平变化

Study of Cyclic Amp in Cardiomyocytes of Heart Failure Rat which Transfected Ad5-ADRbeta2-EGFP by Adenovirus Vector

目的测定重组腺病毒Ad5-ADRbeta2-EGFP转染体外培养心衰大鼠心肌细胞48h后环磷酸腺苷(cAMP)的水平变化。方法重组腺病毒Ad5-ADRbeta2-EGFP转染心衰大鼠心肌细胞,培养48h后,检测心肌细胞β2-AR蛋白表达和cAMP的水平。结果①心肌细胞β2-AR蛋白表达测定:心衰组与对照组(假手术组)相比没有明显差异(P>0.05),心衰+β2组与心衰组、对照组相比均明显升高(P<0.05)。②心肌细胞cAMP水平:心衰组与对照组(假手术组)相比明显减少(P<0.01),心衰+β2组与心衰组相比明显升高(P<0.01),心衰+ISO组与心衰组相比明显升高(P<0.01),心衰+ISO+β2组与心衰组相比明显升高(P<0.01),心衰+ISO+β2组与心衰+β2组、心衰+ISO组相比没有明显差异(P>0.05)。结论重组腺病毒Ad5-ADRbeta2-EGFP转染体外培养心衰大鼠心肌细胞后,β2-AR蛋白表达上调,同时伴有基础cAMP逆转恢复正常水平,其可能与β2-AR活性的内在活化有关。

Objective To investigate the level of cyclic AMP（cAMP） in cardiomyocytes from heart failure rat which transfected AdS-ADRbeta2-EGFP by adenovirus vector. Methods The expression of β2-AR and level of cAMP on heart failure rat ventricular myocytes which transfected AdS-ADRbeta2-EGFP by adenovirus vector and cultured 48 hours. Results （1） Western blot demonstrated the presence of β2-AR protein has no significance between the heart failure group and the control group （ P 〉 0.05）, but the presence of β2-AR protein of the heart failure ＋β2 group was significantly higher than that of the control group and the heart failure group（ P 〈 0.05 ）. （2） ELISA demonstrated the level of cAMP in the heart failure group was significantly lower than that of the control group（P 〈0.01 ）. That of the heart failure ＋ β2 group was significantly higher than the heart failure group（P 〈0.01 ）. But the statistical analysis on the level of cAMP has no significance in the heart failure ＋ ISO ＋ β2 group,the heart failure ＋ β2 group and the heart failure group （P 〉 0.05）. Conclusion After the β2-AR cDNA was transfected 48 hours in vitro to the heart failure rat cardiomyocytes, the presence of β2-AR protein increased and the level of cAMP back to normal. The result suggested that β2-adrenergic receptor activity may be related to the intemal activation.