Nitric oxide induces phosphodiesterase 4B expression in rat pulmonary artery smooth muscle cells
Nitric oxide induces phosphodiesterase 4B expression in rat pulmonary artery smooth muscle cells
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DOI：
10.1152/ajplung.00298.2005
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发表时间：
2006-04-01
影响因子：
4.9
通讯作者：
Bloch, KD
中科院分区：
文献类型：
--
作者：
Busch, CJ;Liu, HL;Bloch, KD
磷酸二酯酶（PDE）代谢环核苷酸，限制血管扩张剂如前列环素和一氧化氮（NO）的作用。本研究应用基因芯片技术研究了NO对大鼠肺动脉平滑肌细胞PDE基因表达的影响。用S-亚硝基-L-谷胱甘肽（GSNO）孵育rPASMC以剂量和时间依赖性方式增加特异性代谢cAMP（PDE 4 B）的PDE亚型的表达。GSNO增加PDE 4 B蛋白水平，并且在GSNO处理的rPASMC中，咯利普兰可降解的PDE活性比未处理的细胞高2.3 +/-1.0倍。可溶性鸟苷酸环化酶（sGC）抑制剂1H-[1，2，4]恶二唑并[4，3，-a]喹喔啉-1-酮和cAMP依赖性蛋白激酶抑制剂H89可阻止GSNO诱导PDE 4 B基因表达，但蛋白激酶G（PKG）抑制剂Rp-8-pCPT-cGMP和KT-5823则不能。IL-1 β和肿瘤坏死因子-α共同孵育rPASMC可诱导PDE 4 B基因表达，这种作用可被NO合成酶2（NOS 2）拮抗剂L-N-6-（1-亚氨基乙基）赖氨酸抑制。GSNO诱导的PDE 4 B mRNA水平的增加被放线菌素D阻断，但被放线菌酮增强。用特异性显性负性cAMP反应元件结合蛋白（CREB）突变体的腺病毒感染rPASMC抑制GSNO诱导的PDE 4 B基因表达增加。这些结果表明，暴露于NO的rPASMC诱导PDE 4 B的表达，通过一种机制，需要cGMP合成的sGC，而不是PKG。GSNO诱导的PDE 4 B基因表达的增加是CREB依赖性的。这些发现表明，NO增加cAMP特异性PDE的表达，并为cGMP和cAMP信号通路之间的新型“串扰”机制提供证据。
Phosphodiesterases (PDE) metabolize cyclic nucleotides limiting the effects of vasodilators such as prostacyclin and nitric oxide ( NO). In this study, DNA microarray techniques were used to assess the impact of NO on expression of PDE genes in rat pulmonary arterial smooth muscle cells (rPASMC). Incubation of rPASMC with S-nitroso-L-glutathione (GSNO) increased expression of a PDE isoform that specifically metabolizes cAMP (PDE4B) in a dose- and time-dependent manner. GSNO increased PDE4B protein levels, and rolipram-inhibitable PDE activity was 2.3 +/- 1.0-fold greater in GSNO-treated rPASMC than in untreated cells. The soluble guanylate cyclase (sGC) inhibitor, 1H-[1,2,4] oxadiazolo[4,3,-a]quinoxalin-1-one, and the cAMP-dependent protein kinase inhibitor, H89, prevented induction of PDE4B gene expression by GSNO, but the protein kinase G (PKG) inhibitors, Rp-8-pCPT-cGMPs and KT-5823, did not. Incubation of rPASMC with IL-1 beta and tumor necrosis factor-alpha induced PDE4B gene expression, an effect that was inhibited by L-N-6-(1-iminoethyl)lysine, an antagonist of NO synthase 2 (NOS2). The GSNO-induced increase in PDE4B mRNA levels was blocked by actinomycin D but augmented by cycloheximide. Infection of rPASMC with an adenovirus specifying a dominant negative cAMP response element binding protein ( CREB) mutant inhibited the GSNO-induced increase of PDE4B gene expression. These results suggest that exposure of rPASMC to NO induces expression of PDE4B via a mechanism that requires cGMP synthesis by sGC but not PKG. The GSNO-induced increase of PDE4B gene expression is CREB dependent. These findings demonstrate that NO increases expression of a cAMP-specific PDE and provide evidence for a novel "cross talk" mechanism between cGMP and cAMP signaling pathways.