Glennon P E, Kaddoura S, Sale E M, Sale G J, Fuller S J, Sugden P H
National Heart and Lung Institute, Imperial College of Science, Technology and Medicine, London, UK.
Circ Res. 1996 Jun;78(6):954-61. doi: 10.1161/01.res.78.6.954.
An antisense oligodeoxynucleotide (ODN) approach was used to investigate whether mitogen-activated protein kinase (MAPK) is necessary for the hypertrophic response in cardiac myocytes. A phosphorothioate-protected 17-mer directed against the initiation of translation sites of the p42 and p44 MAPK isoform mRNAs was introduced into cultured cardiac myocytes by liposomal transfection. At an antisense ODN concentration of 0.2 mumol/L, p42 MAPK protein was reduced by 82% (immunoblot) after 48 hours, and p42 and p44 MAPK activities were reduced by 44% and 60%, respectively. The same concentration of anti-MAPK ODN inhibited development of the morphological features of hypertrophy (sarcomerogenesis, increased cell size) in myocytes exposed to phenylephrine. Phenylephrine-induced activation of the atrial natriuretic factor (ANF) promoter (measured by the activity of a transfected ANF promoter/luciferase reporter gene) and induction of ANF mRNA (measured by RNase protection assay) were also attenuated. We conclude that MAPK is important for the development of the hypertrophic phenotype in this model of hypertrophy.
采用反义寡脱氧核苷酸（ODN）方法来研究丝裂原活化蛋白激酶（MAPK）对于心肌细胞肥大反应是否必要。通过脂质体转染将一种针对p42和p44 MAPK亚型mRNA翻译起始位点的硫代磷酸酯保护的17聚体引入培养的心肌细胞。在反义ODN浓度为0.2 μmol/L时，48小时后p42 MAPK蛋白减少了82%（免疫印迹法），p42和p44 MAPK活性分别降低了44%和60%。相同浓度的抗MAPK ODN抑制了暴露于去甲肾上腺素的心肌细胞肥大形态特征（肌节形成、细胞大小增加）的发展。去甲肾上腺素诱导的心房利钠因子（ANF）启动子激活（通过转染的ANF启动子/荧光素酶报告基因的活性来测量）和ANF mRNA诱导（通过核糖核酸酶保护测定法测量）也减弱了。我们得出结论，在这种肥大模型中，MAPK对于肥大表型的发展很重要。
