Nishimura K, Tamaoki J, Aoshiba K, Isono K, Nagai A
First Department of Medicine, Tokyo Women's Medical College, Japan.
Nihon Kokyuki Gakkai Zasshi. 1998 May;36(5):428-32.
To determine whether stimulation of beta-adrenoceptors affects proliferation of airway epithelial cells and, if so, whether activation of mitogen-activated protein kinase (MAPK) is involved, we studied cultured human bronchial epithelial (16-HBE) cells in vitro. The 16-HBE cells were grown to subconfluence in 96-well plates, and their growth was inhibited by incubation in serum-free medium for 72 h. The cells were ten incubated in the presence of saltbutamol (SAL, 10(-7) M), a specific beta(2)-adrenoceptor agonist. Proliferation of the cells was evaluated by MTT assay and total DNA content, and activation of MAPK was assessed by immunocytochemistry and Western blotting for phosphorylated MAPK (phospho-MAPK). Immunocytochemistry and immunoblots demonstrated that phospho-MAPK was expressed within minutes of SAL exposure. This effect of SAL was as potent as that of 10% serum, and was greatly inhibited by treatment with propranolol. These results suggest that SAL is a potent mitogen of airway epithelial cells and that its effect may be exerted by beta(2)-adrenocepter-mediated activation of MAPK.
为了确定β-肾上腺素能受体的刺激是否会影响气道上皮细胞的增殖，以及如果有影响，丝裂原活化蛋白激酶（MAPK）的激活是否参与其中，我们在体外研究了培养的人支气管上皮（16-HBE）细胞。将16-HBE细胞在96孔板中培养至亚汇合状态，然后在无血清培养基中孵育72小时以抑制其生长。接着将细胞在特异性β₂-肾上腺素能受体激动剂沙丁胺醇（SAL，10⁻⁷ M）存在的情况下孵育。通过MTT法和总DNA含量评估细胞增殖，并通过免疫细胞化学和针对磷酸化MAPK（磷酸化-MAPK）的蛋白质印迹法评估MAPK的激活。免疫细胞化学和免疫印迹表明，在暴露于SAL的几分钟内就有磷酸化-MAPK表达。SAL的这种作用与10%血清的作用一样有效，并且用普萘洛尔处理可大大抑制这种作用。这些结果表明，SAL是气道上皮细胞的一种有效促有丝分裂原，其作用可能通过β₂-肾上腺素能受体介导的MAPK激活来发挥。
