Selective blockade of phosphodiesterase types 2, 5 and 9 results in cyclic 3′5′ guanosine monophosphate accumulation in retinal pigment epithelium cells
Selective blockade of phosphodiesterase types 2, 5 and 9 results in cyclic 3′5′ guanosine monophosphate accumulation in retinal pigment epithelium cells
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DOI：
10.1136/bjo.2006.100628
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发表时间：
2007-03-01
影响因子：
4.1
通讯作者：
de Vente, J.
中科院分区：
文献类型：
--
作者：
Diederen, R. M. H.;La Heij, E. C.;de Vente, J.
目的：研究哪种磷酸二酯酶（PDE）参与了视网膜色素上皮（RPE）细胞中的环磷酸3959鸟苷单磷酸的分解。方法：培养的RPE细胞（D407细胞系）中的CGMP含量，在非Immunocyocyto Chemistry中评估 - 选择性或同工型选择性PDE抑制剂组合与颗粒的鸟叶基环化酶刺激剂房屋纳特里肽（ANP）或可溶性鸟叶尼环酶刺激剂硝化钠（SNP）一起使用。使用非放射性原位杂交化研究了PDE2，PDE5和PDE9的mRNA表达在培养的人RPE细胞和大鼠RPE细胞层中进行了研究：分泌：在没有PDE抑制剂的情况下，培养的RPE细胞中CGMP水平的CGMP水平非常低。在与Bay60-7550孵育后，在培养的人RPE细胞中很容易检测CGMP积累，作为选择性PDE2抑制剂，西地那非作为选择性PDE5抑制剂或SCH51866作为选择性PDE9抑制剂。在存在PDE抑制的情况下，刺激颗粒鸟叶烷环酶后，CGMP含量显着增加。在所有培养的人RPE细胞以及大鼠RPE细胞层中都检测到PDE2，PDE5和PDE9的mRNA。结论：PDE2，PDE5和PDE9在RPE细胞的CGMP代谢中均起作用。
Aim: To investigate which phosphodiesterase (PDE) is involved in regulating cyclic 3959 guanosine monophosphate breakdown in retinal pigment epithelium (RPE) cells.Methods: cGMP content in the cultured RPE cells (D407 cell line) was evaluated by immunocytochemistry in the presence of non-selective or isoform-selective PDE inhibitors in combination with the particulate guanylyl cyclase stimulator atrial natriuretic peptide (ANP) or the soluble guanylyl cyclase stimulator sodium nitroprusside (SNP). mRNA expression of PDE2, PDE5 and PDE9 was studied in cultured human RPE cells and rat RPE cell layers using non-radioactive in situ hybridisation.Results: In the absence of PDE inhibitors, cGMP levels in cultured RPE cells are very low. cGMP accumulation was readily detected in cultured human RPE cells after incubation with Bay60-7550 as a selective PDE2 inhibitor, sildenafil as a selective PDE5 inhibitor or Sch51866 as a selective PDE9 inhibitor. In the presence of PDE inhibition, cGMP content increased markedly after stimulation of the particulate guanylyl cyclase. mRNA of PDE2, PDE5 and PDE9 was detected in all cultured human RPE cells and also in rat RPE cell layers.Conclusions: PDE2, PDE5 and PDE9 have a role in cGMP metabolism in RPE cells.