Yau L, Zahradka P
Institute of Cardiovascular Sciences, St. Boniface General Hospital Research Centre, University of Manitoba, Winnipeg, Canada.
Mol Cell Biochem. 1997 Jul;172(1-2):59-66.
Mitogen-activated protein kinase (MAPK) is a key modulator of cytoplasmic-nuclear signal transmission, and for this reason measurement of MAPK activity has become very popular. Monitoring of MAPK activity may be particularly relevant to the cardiovascular system where it has already been shown that the stimulation of cardiomyocytes and smooth muscle cells by stretch and by growth factors activates MAPK. Since both growth factors and mechanical stress are causal agents for certain pathologies, enhanced MAPK activity may be a good predictor of disease progression. A variety of methods have been designed to measure the activation of this enzyme including an in vitro assay coupled to either gel electrophoresis or binding to P81 paper, an activity gel assay to detect p42/44 isoforms and, more recently, monitoring MAPK phosphorylation using immunoblot detection. The validity of the latter method is based on the correlation between MAPK activity and the degree of phosphorylation. The antibodies have also been of use in the detection of MAPK translocation in cell monolayers. In this report, we discuss the advantages and disadvantages of all MAPK detection methods and demonstrate an additional application for the MAPK antibodies using an in vitro restenosis model. In addition, the utility of MAPK measurements to smooth muscle pathophysiology and vascular injury (as a predictor of injury) has been assessed.
丝裂原活化蛋白激酶（MAPK）是细胞质-细胞核信号传导的关键调节因子，因此，对MAPK活性的检测已变得非常普遍。监测MAPK活性可能与心血管系统特别相关，在心血管系统中，已经表明拉伸和生长因子对心肌细胞和平滑肌细胞的刺激会激活MAPK。由于生长因子和机械应激都是某些病理状态的致病因素，增强的MAPK活性可能是疾病进展的良好预测指标。已经设计了多种方法来测量这种酶的激活，包括与凝胶电泳或与P81纸结合的体外测定、检测p42/44亚型的活性凝胶测定，以及最近使用免疫印迹检测监测MAPK磷酸化。后一种方法的有效性基于MAPK活性与磷酸化程度之间的相关性。这些抗体也已用于检测细胞单层中的MAPK易位。在本报告中，我们讨论了所有MAPK检测方法的优缺点，并使用体外再狭窄模型展示了MAPK抗体的另一种应用。此外，还评估了MAPK测量对平滑肌病理生理学和血管损伤（作为损伤预测指标）的实用性。
