论文部分内容阅读
目的 观察血管紧张素(Ang)Ⅱ作用心肌细胞后细胞外信号调节激酶 (ERK)活化、核转位情况、c fos基因表达及在受体和信号ERK水平进行干预的影响。方法 采用原代培养的心肌细胞,分别从细胞膜受体和细胞内信号ERK层面进行干预,用细胞免疫化学观察磷酸化ERK入核过程,RT PCR测定c fos基因表达量。结果 AngⅡ刺激心肌细胞可见胞核内出现磷酸化ERK染色,Valsartan(10-5mol/L)、PD98059 ( 5×10-5mol/L)可阻断AngⅡ引起的ERK活化、入核过程,而CGP42112A(10-5mol/L)则无阻断作用;AngⅡ可促进心肌细胞c fosmRNA表达,在 30~60min达高峰,Valsartan、PD98059均可抑制该作用,CGP42112A对该作用无明显影响。结论 AngⅡ可磷酸化胞质ERK,并使其发生转位入核,ERK的跨核转运可能是AngⅡ诱导c fos原癌基因表达的前提。
Objective To observe the effects of angiotensin Ⅱ on the activation of extracellular signal - regulated kinase (ERK), nuclear translocation, c fos gene expression and the intervention of ERK at receptor and signal. Methods Primary cultured cardiomyocytes were used to intervene respectively at the cell membrane receptor and the intracellular signal ERK. The phosphorylation of ERK was detected by immunocytochemistry and the expression of c fos gene was detected by RT PCR. Results AngⅡ stimulated cardiomyocytes showed phosphorylation of ERK staining, Valsartan (10-5mol / L), PD98059 (5 × 10-5mol / L) can be blocked by Ang Ⅱ-induced ERK activation, nuclear process, and CGP42112A 10-5mol / L) did not block the effect; Ang Ⅱ can promote cardiac myocyte c fosmRNA expression peaked at 30 ~ 60min, Valsartan, PD98059 can inhibit this effect, CGP42112A no significant effect on this effect. Conclusion AngⅡ phosphorylates ERK and translocates into the nucleus. The transnational nuclear translocation of ERK may be the prerequisite for the induction of c fos proto-oncogene by AngⅡ.