Expression and activity of cGMP-dependent phosphodiesterases is up-regulated by lipopolysaccharide (LPS) in rat peritoneal macrophages
Expression and activity of cGMP-dependent phosphodiesterases is up-regulated by lipopolysaccharide (LPS) in rat peritoneal macrophages
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DOI：
10.1016/j.bbamcr.2006.10.008
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发表时间：
2007-02-01
影响因子：
5.1
通讯作者：
Gorczyca, Wojciech A.
中科院分区：
文献类型：
--
作者：
Witwicka, Hanna;Kobialka, Marcin;Gorczyca, Wojciech A.
环鸟苷酸（cGMP）调节巨噬细胞的炎症反应，但其潜在的分子机制仍然知之甚少。为了寻找大鼠腹腔巨噬细胞中可能受cGMP调控的蛋白质，本研究分析了cGMP水解和cGMP调控的磷酸二酯酶（PDE）的表达和活性。发现新鲜分离的腹膜渗出液巨噬细胞（PEM）表达属于家族PDE 1 -3、PDE 5、PDE 10和PDE 11的酶。底物特异性、对抑制剂的敏感性和亚细胞定位的分析表明，PDE 2和PDE 3是PEM中主要的cGMP调节的PDE亚型。PDE表达的概况通过保持PEM在培养物中和用细菌内毒素（LPS）处理而改变。24 μ l培养后，PDE 5不存在，PDE 2、PDE 3和PDE II的水平显著降低。然而，它们的表达和活性在用LPS处理培养的细胞后恢复。观察到TNF α表达的类似变化模式，但未观察到鸟苷酸环化酶A（GC-A）。LPS上调PDE的表达也在居民腹膜巨噬细胞（RPM），虽然不是所有的PDE存在于PEM中检测到RPM。总之，我们的研究结果表明，在大鼠PMs表达cGMP依赖的PDE与细胞的活化状态呈正相关。此外，这些PDE中的大多数也水解cAMP的事实表明，cGMP可以在巨噬细胞中发挥cAMP信号传导的有效调节剂的作用。(c)2006 Elsevier B. V.保留所有权利。
It has been shown that cyclic GMP (cGMP) modulates the inflammatory responses of macrophages, but the underlying molecular mechanisms are still poorly understood. Looking for proteins potentially regulated by cGMP in rat peritoneal macrophages (PMs), in this study we analyzed expression and activity of cGMP-hydrolyzing and cGMP-regulated phosphodiesterases (PDEs). It was found that freshly isolated peritoneal exudate macrophages (PEMs) express enzymes belonging to families PDE1-3, PDE5, PDE10, and PDE11. Analysis of substrate specificity, sensitivity to inhibitors, and subcellular localization showed that PDE2 and PDE3 are the main cGMP-regulated PDE isoforms in PEMs. The profile of PDE expression was altered by maintaining PEMs in culture and treatment with bacterial endotoxin (LPS). After 24 It culture, PDE5 was not present and the levels of PDE2, PDE3, and PDE I I were markedly decreased. However, their expression and activity was recovered after treatment of cultured cells with LPS. A similar pattern of changes was observed for the expression of TNF alpha, but not for guanylyl cyclase A (GC-A). LPS up-regulated PDE expression also in resident peritoneal macrophages (RPMs), although not all PDEs present in PEMs were detected in RPMs. Taken together, our results show that in rat PMs expression of cGMP-dependent PDEs positively correlates with the activation state of cells. Moreover, the fact that most of these PDEs hydrolyze also cAMP indicates that cGMP can play a role of potent regulator of cAMP signaling in macrophages. (c) 2006 Elsevier B.V. All rights reserved.