Effect of angiotensin II on the WNK-OSR1/SPAK-NCC phosphorylation cascade in cultured mpkDCT cells and in vivo mouse kidney
Effect of angiotensin II on the WNK-OSR1/SPAK-NCC phosphorylation cascade in cultured mpkDCT cells and in vivo mouse kidney
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DOI：
10.1016/j.bbrc.2010.02.096
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发表时间：
2010-03-19
影响因子：
3.1
通讯作者：
Uchida, Shinichi
中科院分区：
文献类型：
--
作者：
Talati, Gulibaha;Ohta, Akihito;Uchida, Shinichi
在我们最近使用Wnk 4（D561 A/+）敲入小鼠的研究中，我们确定了WNK-OSR 1/SPAK-NaCl协同转运蛋白（NCC）磷酸化级联对于体内调节NCC功能是重要的。NCC的磷酸化是其质膜定位所必需的。先前，血管紧张素II输注显示增加大鼠中NCC的顶端膜表达。因此，我们研究了血管紧张素II是否是培养细胞和体内肾脏中WNK-OSR 1/SPAK-NCC级联反应的上游调节剂。在mpkDCT细胞中，加入血管紧张素II（10（-9）-10（-7）M）后30 min，OSR 1和NCC的磷酸化水平增加，但在18 h后恢复至基线水平。在小鼠中，血管紧张素II（5 ng/g/min）的5分钟输注在注射后30分钟和2小时增加了肾脏中的NCC磷酸化，但在24小时后恢复到基线。这种增加被血管紧张素II受体阻滞剂（缬沙坦）抑制，但不被醛固酮受体阻滞剂（依普利酮）抑制。血管紧张素II（720 ng/天）的10天输注也增加了小鼠肾脏中OSR 1和NCC的磷酸化，缬沙坦和依普利酮都抑制了磷酸化的增加。虽然血管紧张素II被确定为体内WNK-OSR 1/SPAK-NCC级联的上游调节剂，但在血管紧张素II的长期调节中，醛固酮似乎是该信号级联的主要调节剂。(C)2010年爱思唯尔公司All rights reserved.
In our recent study using Wnk4(D561A/+) knockin mice, we determined that the WNK-OSR1/SPAK-NaCl cotransporter (NCC) phosphorylation cascade is important for regulating NCC function in vivo. Phosphorylation of NCC was necessary for its plasma membrane localization. Previously, angiotensin II infusion was shown to increase apical membrane expression of NCC in rats. Therefore, we investigated whether angiotensin II was an upstream regulator for the WNK-OSR1/SPAK-NCC cascade in cultured cells and in vivo kidney. In mpkDCT cells, the phosphorylation of OSR1 and NCC was increased 30 min after the addition of angiotensin II (10(-9)-10(-7) M) but returned to baseline after 18 h. In mice, a 5-min infusion of angiotensin II (5 ng/g/min) increased NCC phosphorylation in the kidney at 30 min and 2 h after the injection but returned to baseline 24 h later. This increase was inhibited by angiotensin II receptor blocker (valsartan) but not by aldosterone receptor blocker (eplerenone). Ten-day infusions of angiotensin II (720 ng/day) also increased phosphorylation of OSR1 and NCC in the mouse kidney, and both valsartan and eplerenone inhibited the increased phosphorylation. Although angiotensin II is identified as an upstream regulator for the WNK-OSR1/SPAK-NCC cascade in vivo, aldosterone appears to be the major regulator of this signal cascade in the long-term regulation by angiotensin II. (C) 2010 Elsevier Inc. All rights reserved.