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缺氧诱导的15-羟二十碳四烯酸(15-hydroxyeicosateteraenoic acid,15-HETE)是引起肺动脉收缩的重要介导因子。15- HETE引起肺动脉收缩的信号转导途径尚不清楚。本研究旨在确定细咆外信号调节激酶1／2(extracellular signal-regulated kinase-1／2, ERK1／2)信号转导通路是否参与15-HETE收缩缺氧大鼠肺动脉的过程。采用组织浴槽肺动脉环张力检测、蛋白质免疫印迹 (Western blot)和免疫细胞化学方法。制备缺氧大鼠动物模型,成年雄性Wistar大鼠在低氧环境下(吸入氧分数为0．12)正常喂养 9 d。显微分离直径1～1．5mm肺动脉,剪成长为3 mm的动脉环,进行血管张力检测。用ERK1／2 上游激酶(MEK)抑制剂PD98059 抑制ERK1／2活性。结果显示,PD98059可明显抑制15-HETE对缺氧大鼠肺动脉环的收缩作用。在去除内皮的肺动脉环, PD98059仍可明显降低15-HETE的缩血管作用。Western blot和免疫细胞化学结果都显示,15-HETE能促进ERK1／2磷酸化。 由此表明ERK1/2信号转导通路参与15-HETE收缩缺氧大鼠肺动脉的过程。
Hypoxia-induced 15-hydroxyeicosateteraenoic acid (15-HETE) is an important mediator of pulmonary artery contraction. The signaling pathways by which 15-HETE causes pulmonary artery contraction are not yet known. The purpose of this study was to determine whether extracellular signal-regulated kinase-1 (ERK1 / 2) signal transduction pathway is involved in the process of 15-HETE contraction in rat pulmonary arteries. Tissue bath pulmonary artery tension testing, Western blot and immunocytochemistry were used. Animal models of hypoxia were established. Adult male Wistar rats were fed for 9 days under hypoxia (oxygen fraction 0.12). Microscopic separation of pulmonary artery diameter 1 ~ 1.5mm, 3mm length of the growth of the cutaneous artery, vascular tension test. Inhibition of ERK1 / 2 activity with ERK1 / 2 upstream kinase (MEK) inhibitor PD98059. The results showed that PD98059 could significantly inhibit the contraction of pulmonary artery rings induced by 15-HETE in hypoxic rats. In removing the endothelial pulmonary artery rings, PD98059 still significantly reduced the vasoconstriction of 15-HETE. Western blot and immunocytochemistry showed that 15-HETE promoted ERK1 / 2 phosphorylation. Thus ERK1 / 2 signal transduction pathway involved in the process of 15-HETE contraction of rat pulmonary artery.