Department of Physiology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand.
Biomed Res Int. 2013;2013:346480. doi: 10.1155/2013/346480. Epub 2013 Jan 22.
Previous in vitro studies indicated that aldosterone nongenomically phosphorylates epidermal growth factor receptor (EGFR) through activation of upstream signals, heat shock protein 90 β (Hsp90 β ), and cytosolic (c)-Src kinase. We demonstrated that aldosterone rapidly elevates EGFR phosphorylation in rat kidney. There are no in vivo data regarding renal Hsp90( α and β ) and phosphorylated (p)c-Src protein expressions. The present study further investigates the expressions of these proteins. Male Wistar rats were intraperitoneally injected with normal saline solution or aldosterone (Aldo: 150 μ g/kg BW). After 30 minutes, abundances and localizations of these proteins were determined. Aldosterone enhanced renal Hsp90 β protein abundance (P < 0.001), but Hsp90 α and pc-Src protein levels remained unaltered. Expression of Hsp90( α and β ) was induced prominently in the proximal convoluted tubules (PCTs). Activation of Hsp90 α was observed in vascular and outer medulla regions, whereas Hsp90 β was induced in the cortex. Immunoreactivity of pc-Src was elevated in PCT with obvious staining at the luminal membrane. This in vivo study is the first to demonstrate that aldosterone nongenomically elevates Hsp90( α and β ) protein expressions in rat kidney. Aldosterone had no effect on pc-Src protein levels but modulated localization. These results indicate that aldosterone regulates upstream mediators of EGFR transactivation in vivo.
先前的体外研究表明,醛固酮通过激活上游信号、热休克蛋白 90β(Hsp90β)和细胞质(c)-Src 激酶非基因组地磷酸化表皮生长因子受体(EGFR)。我们证明醛固酮可迅速增加大鼠肾脏中 EGFR 的磷酸化。关于肾脏 Hsp90(α和β)和磷酸化(p)c-Src 蛋白表达,尚无体内数据。本研究进一步研究了这些蛋白的表达。雄性 Wistar 大鼠腹膜内注射生理盐水或醛固酮(Aldo:150μg/kgBW)。30 分钟后,测定这些蛋白的丰度和定位。醛固酮增强了肾脏 Hsp90β 蛋白的丰度(P<0.001),但 Hsp90α和 pc-Src 蛋白水平保持不变。Hsp90(α和β)的表达在近端曲管(PCTs)中明显诱导。在血管和外髓质区域观察到 Hsp90α的激活,而 Hsp90β在皮质中被诱导。pc-Src 的免疫反应性在 PCT 中升高,在腔膜处有明显染色。这项体内研究首次证明,醛固酮非基因组地增加大鼠肾脏中 Hsp90(α和β)的蛋白表达。醛固酮对 pc-Src 蛋白水平没有影响,但调节了定位。这些结果表明,醛固酮在体内调节 EGFR 转激活的上游介质。