Molecular mechanisms of angiotensin II stimulation on aquaporin-2 expression and trafficking
Molecular mechanisms of angiotensin II stimulation on aquaporin-2 expression and trafficking
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DOI：
10.1152/ajprenal.00469.2010
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发表时间：
2011-05-01
影响因子：
4.2
通讯作者：
Schrier, Robert W.
中科院分区：
文献类型：
--
作者：
Li, Chunling;Wang, Weidong;Schrier, Robert W.
李超，王伟，riward CJ, Lanaspa MA, Summer S, Schrier RW。血管紧张素II刺激水通道蛋白-2表达和转运的分子机制。[J] .中国生物医学工程学报，2016,31(5):555 - 561。2011年2月16日首次出版；doi: 10.1152 / ajprenal.00469.2010。-ANG II在肾脏水钠调节中起重要作用。在永生化小鼠肾收集管主细胞（mpkCCD(cl4)）细胞系中，我们用ANG II处理细胞，并通过免疫印迹、免疫荧光和RT-PCR检测水通道蛋白-2 （AQP2）蛋白的表达、转运和mRNA水平。孵育24 h后，在浓度为10(-10)M时观察到ANG ii诱导的AQP2蛋白表达，并呈剂量依赖性增加。ANG II （10(-7) M）在0.5、1、2、6和24 h时增加AQP2蛋白表达和mRNA水平。免疫荧光研究显示，ANG II在30 min至6 h时增加了AQP2的顶膜靶向。接下来，研究ANG II诱导AQP2表达的信号通路。PKC抑制剂Ro 31-8220 （5 × 10(-6) M）和PKA抑制剂H89 （10(-5) M）分别阻断ANG ii诱导的AQP2表达。钙调素抑制剂W-7显著降低ANG ii和/或ddavp刺激的AQP2表达。ANG II （10(-9) M）和/或dDAVP （10(-10) M）刺激AQP2蛋白水平和cAMP积累，通过加压素V2受体（V2R）拮抗剂SR121463B （10(-8) M）预处理完全阻断。血管紧张素AT(1)受体（AT1R）拮抗剂氯沙坦（3 × 10(-6) M）预处理可阻断ANG II （10(-9) M）诱导的AQP2蛋白表达和cAMP积累，部分阻断dDAVP (10(-10) M)-和dDAVP + ANG II诱导的AQP2蛋白表达和cAMP积累。综上所述，ANG II调节肾集管主细胞中AQP2蛋白、转运和基因表达。ANG ii诱导的AQP2表达通过V2和AT(1)受体参与cAMP、PKC、PKA和钙调素信号通路。
Li C, Wang W, Rivard CJ, Lanaspa MA, Summer S, Schrier RW. Molecular mechanisms of angiotensin II stimulation on aquaporin-2 expression and trafficking. Am J Physiol Renal Physiol 300: F1255-F1261, 2011. First published February 16, 2011; doi:10.1152/ajprenal.00469.2010.-ANG II plays a major role in renal water and sodium regulation. In the immortalized mouse renal collecting duct principal cells (mpkCCD(cl4)) cell line, we treated cells with ANG II and examined aquaporin-2 (AQP2) protein expression, trafficking, and mRNA levels, by immunoblotting, immunofluorescence, and RT-PCR. After 24-h incubation, ANG II-induced AQP2 protein expression was observed at the concentration of 10(-10) M and increased in a dose-dependent manner. ANG II (10(-7) M) increased AQP2 protein expression and mRNA levels at 0.5, 1, 2, 6, and 24 h. Immunofluorescence studies showed that ANG II increased the apical membrane targeting of AQP2 from 30 min to 6 h. Next, the signaling pathways underlying the ANG II-induced AQP2 expression were investigated. The PKC inhibitor Ro 31-8220 (5 X 10(-6) M) and the PKA inhibitor H89 (10(-5) M) blocked ANG II-induced AQP2 expression, respectively. Calmodulin inhibitor W-7 markedly reduced ANG II-and/or dDAVP-stimulated AQP2 expression. ANG II (10(-9) M) and/or dDAVP (10(-10) M) stimulated AQP2 protein levels and cAMP accumulation, which was completely blocked by pretreatment with the vasopressin V2 receptor (V2R) antagonist SR121463B (10(-8) M). Pretreatment with the angiotensin AT(1) receptor (AT1R) antagonist losartan (3 X 10(-6) M) blocked ANG II (10(-9) M)-stimulated AQP2 protein expression and cAMP accumulation, and partially blocked dDAVP (10(-10) M)- and dDAVP + ANG II-induced AQP2 protein expression and cAMP accumulation. In conclusion, ANG II regulates AQP2 protein, trafficking, and gene expression in renal collecting duct principal cells. ANG II-induced AQP2 expression involves cAMP, PKC, PKA, and calmodulin signaling pathways via V2 and AT(1) receptors.