抗利尿激素抑制马-达二氏犬肾细胞中表皮生长因子刺激的丝裂原活化蛋白激酶级联反应。

AVP inhibits EGF-stimulated MAP kinase cascade in Madin-Darby canine kidney cells.

作者信息

Yamada T, Terada Y, Homma M K, Nonoguchi H, Sasaki S, Yuasa Y, Tomita K, Marumo F

机构信息

Second Department of Internal Medicine, Tokyo Medical and Dental University, Japan.

出版信息

Kidney Int. 1995 Sep;48(3):745-52. doi: 10.1038/ki.1995.346.

DOI:10.1038/ki.1995.346

PMID:7474660

Abstract

We investigated the effects of epidermal growth factor (EGF) and arginine vasopressin (AVP) on Raf-1-MAP kinase cascade, including Raf-1-kinase (Raf-1-K), MAP kinase kinase (MAPKK), MAP kinase (MAPK) and S6 kinase (S6K) in Madin-Darby canine kidney (MDCK) cells. In a dose-dependent manner (10(-10) M to 10(-6) M), EGF increased autophosphorylation of Raf-1-K and activated MAPKK, MAPK and S6K. Sequential activation of these kinases was indicated by their peak times of activation (Raf-1-K 5 min; MAPKK 10 min; MAPK 15 min; and S6K 30 min). AVP (10(-9) M to 10(-6) M) inhibited EGF-stimulated MAP kinase cascade. 8-Bromo-cyclic AMP (cAMP) could mimic the inhibitory effect of AVP on EGF-stimulated MAP kinase cascade. These results were confirmed using H-89, an inhibitor of protein kinase A (PKA) that blocked the effect of AVP on EGF-stimulated MAPK activity. We conclude that AVP inhibits EGF-stimulated Raf-1-K, MAPKK, MAPK, and S6K activity via cAMP in MDCK cells. Our results indicate that MAP kinase cascade may play an important role in integrating the effects of AVP and EGF on distal tubule function.

摘要

我们研究了表皮生长因子（EGF）和精氨酸加压素（AVP）对Madin-Darby犬肾（MDCK）细胞中Raf-1-丝裂原活化蛋白激酶（MAP）激酶级联反应的影响，该级联反应包括Raf-1激酶（Raf-1-K）、MAP激酶激酶（MAPKK）、MAP激酶（MAPK）和S6激酶（S6K）。EGF以剂量依赖方式（10⁻¹⁰ M至10⁻⁶ M）增加Raf-1-K的自磷酸化，并激活MAPKK、MAPK和S6K。这些激酶的顺序激活通过其激活的峰值时间表明（Raf-1-K 5分钟；MAPKK 10分钟；MAPK 15分钟；S6K 30分钟）。AVP（10⁻⁹ M至10⁻⁶ M）抑制EGF刺激的MAP激酶级联反应。8-溴环磷酸腺苷（cAMP）可模拟AVP对EGF刺激的MAP激酶级联反应的抑制作用。使用蛋白激酶A（PKA）抑制剂H-89证实了这些结果，H-89阻断了AVP对EGF刺激的MAPK活性的影响。我们得出结论，在MDCK细胞中，AVP通过cAMP抑制EGF刺激的Raf-1-K、MAPKK、MAPK和S6K活性。我们的结果表明，MAP激酶级联反应可能在整合AVP和EGF对远端小管功能的影响中起重要作用。