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目的有报道表明staurosporine作为一种蛋白激酶C(PKC)的抑制剂,可以在多种细胞系内调节细胞外信号调节激酶(ERK1/2)的活性,但是它的衍生物staurosporine agly-cone(SA)是否具有相同的作用还不清楚,本次实验旨在阐明其对ERK1/2活性的调节作用。方法培养至4~8代的大鼠肺动脉平滑肌细胞,经过SA处理后提取细胞蛋白,应用Western blot方法检测磷酸化ERK1/2的含量,观察不同浓度和时间点的SA对ERK1/2活性的影响。结果在纳摩尔浓度水平的SA可以降低大鼠肺动脉平滑肌细胞中ERK1/2的磷酸化水平,但是30μmol.L-1的SA可以激活ERK1/2,这种激活作用可以被丝裂素活化蛋白激酶的激酶(MEK)的抑制剂PD98059或蛋白激酶A(PKA)的激活剂异丙肾上腺素(isoproterenol)所抑制。结论SA对肺动脉平滑肌细胞中的ERK1/2活性有双向调节作用,这种作用是通过PKC和(或)PKA通路产生的。
It has been reported that staurosporine, as an inhibitor of protein kinase C (PKC), regulates the activity of extracellular signal-regulated kinase (ERK1 / 2) in a variety of cell lines, but its derivative staurosporine agly-cone ) Whether it has the same effect is unclear, this experiment aims to clarify its regulation of ERK1 / 2 activity. Methods Cultured rat pulmonary artery smooth muscle cells from 4 to 8 passages were cultured and the cell proteins were extracted after SA treatment. The content of phosphorylated ERK1 / 2 was detected by Western blot and the effects of SA at different concentrations and time points on the activity of ERK1 / 2 . Results SA at nanomolar level could decrease the phosphorylation of ERK1 / 2 in rat pulmonary arterial smooth muscle cells, but SA at 30μmol.L-1 could activate ERK1 / 2, and this activation could be inhibited by mitogen-activated protein kinase (MEK) inhibitor PD98059 or an inhibitor of protein kinase A (PKA) isoproterenol. Conclusions SA exerts bidirectional modulation of ERK1 / 2 activity in pulmonary artery smooth muscle cells through PKC and / or PKA pathways.