2-methoxyestradiol binding of GPR30 down-regulates angiotensin AT(1) receptor.
2-methoxyestradiol binding of GPR30 down-regulates angiotensin AT(1) receptor.
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DOI：
10.1016/j.ejphar.2013.10.064
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发表时间：
2014-01-15
影响因子：
5
通讯作者：
Thekkumkara, Thomas
中科院分区：
文献类型：
--
作者：
Koganti, Sivaramakrishna;Snyder, Russell;Gumaste, Upendra;Karamyan, Vardan T.;Thekkumkara, Thomas
关键词：
控制血管紧张素 AT1 受体功能已被证明对许多病理生理疾病具有保护作用。尽管雌激素代谢物 2-甲氧基雌二醇 (2ME2) 可以独立于核受体下调血管紧张素 AT1 受体的表达，但尚未确定特定的细胞靶标。本研究的重点是在连续传代的大鼠肝上皮细胞系中鉴定和验证负责 2ME2 介导的血管紧张素 AT1 受体下调的细胞靶标。分离细胞膜并用于测定2ME2特异性结合。暴露于[3H]2ME2的细胞膜显示出特异性的可饱和结合，这被发现对百日咳毒素(PTx)敏感。在类似条件下，G蛋白偶联受体30 (GPR30)激动剂(G1)和拮抗剂(G15)抑制2ME2特异性结合。在这些细胞中，GPR30 被发现定位于内质网 (ER) 膜。在完整细胞中，G1 下调血管紧张素 AT1 受体的表达，并且这种作用被 G15 逆转。此外，2ME2 介导的表皮生长因子受体 (EGFR) 激活以及随后的 ERK1/2 磷酸化（血管紧张素 AT1 受体下调的重要信号传导步骤）被 G15 消除，表明该信号是 GPR30 依赖性的。此外，还发现 EGF 以 ERK1/2 依赖性方式独立下调血管紧张素 AT1 受体。总之，我们的结果首次证明2ME2对血管紧张素AT1受体的下调依赖于ER膜相关的GRP30。此外，EGFR 的 GPR30 依赖性反式激活和 ERK1/2 磷酸化促进了这种效应。本研究进一步了解 2ME2 的生理意义及其在调节血管紧张素 AT1 受体表达中的作用。
Controlling angiotensin AT1 receptor function has been shown to be protective for many pathophysiological disorders. Although estrogen metabolite, 2-methoxyestradiol (2ME2) can down-regulate angiotensin AT1 receptor expression independently of nuclear receptors, no specific cellular targets have been identified. This study was focused on identification and validation of a cellular target responsible for 2ME2-mediated angiotensin AT1 receptor down-regulation in a continuously passaged rat liver epithelial cell line. Cell membranes were isolated and used to determine 2ME2 specific binding. Cell membranes exposed to [3H]2ME2 showed specific saturable binding, which was found to be pertussis toxin (PTx) sensitive. Under similar conditions, G-protein coupled receptor 30 (GPR30) agonist (G1) and antagonist (G15) inhibited 2ME2 specific binding. In these cells GPR30 was found localized to endoplasmic reticulum (ER) membranes. In intact cells, G1 down-regulated angiotensin AT1 receptor expression and this effect was reversed by G15. Furthermore, 2ME2 mediated activation of epidermal growth factor receptor (EGFR) followed by ERK1/2 phosphorylation, an essential signaling step in angiotensin AT1 receptor down-regulation, was abrogated by G15, suggesting that this signal is GPR30 dependent. Additionally, EGF was found to independently down-regulate angiotensin AT1 receptor in an ERK1/2-dependent manner. In summary, our results demonstrate for the first time that 2ME2 down-regulation of angiotensin AT1 receptor is dependent on ER membrane-associated GRP30. Moreover, this effect is facilitated by GPR30 dependent transactivation of EGFR and ERK1/2 phosphorylation. This study provides further understanding of the physiological significance of 2ME2 and its role in modulating angiotensin AT1 receptor expression.
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DOI：
10.1161/hypertensionaha.110.152298
发表时间：
2010-11
期刊：
Hypertension (Dallas, Tex. : 1979)
影响因子：
--
作者：
Barchiesi F;Lucchinetti E;Zaugg M;Ogunshola OO;Wright M;Meyer M;Rosselli M;Schaufelberger S;Gillespie DG;Jackson EK;Dubey RK
通讯作者：
Dubey RK
影响因子：
4.6
作者：
Kumar, AP;Garcia, GE;Slaga, TJ
通讯作者：
Slaga, TJ
影响因子：
3.4
作者：
Delbeck, Martina;Golz, Stefan;Otto, Christane
通讯作者：
Otto, Christane
影响因子：
11.4
作者：
Gschwind, A;Hart, S;Ullrich, A
通讯作者：
Ullrich, A
影响因子：
3
作者：
Finlay, Trisha M.;Jayanth, Preethi;Szewczuk, Myron R.
通讯作者：
Szewczuk, Myron R.