Nitric oxide-mediated regulation of connexin43 expression and gap junctional intercellular communication in mesangial cells
Nitric oxide-mediated regulation of connexin43 expression and gap junctional intercellular communication in mesangial cells
复制标题
DOI：
10.1681/asn.2004060453
复制
发表时间：
2005-01-01
影响因子：
13.6
通讯作者：
Kitarnura, M
中科院分区：
文献类型：
--
作者：
Jian, Y;Hiramatsu, N;Kitarnura, M
本研究探讨了一氧化氮（NO）在细胞间隙连接通讯（GJIC）调控中的潜在作用。NO供体S-亚硝基-N-乙酰青霉胺（SNAP）孵育系膜细胞（MC）可增强基础和8-溴-cAMP刺激的GJIC以及间隙连接蛋白连接蛋白43（Cx43）的表达。SNAP对Cx4, 3表达的这种增强作用被另外两种NO供体模拟，并被可溶性鸟苷酸环化酶抑制剂1H-[1，2，4]恶二唑并[4，3，-alpha]喹喔啉-1-1显著阻断。鸟苷3 '，5'-环一磷酸（cGMP）类似物8-溴-cGMP产生类似于NO的作用，而另一种cGMP类似物8-pCPT-cGMP选择性激活GMP依赖性激酶而不影响cGMP抑制的磷酸二酯酶（PDE 3），没有作用。此外，NO对Cx43表达的协同作用可被蛋白激酶A抑制剂H89完全阻断，但不能被cGMP依赖性激酶抑制剂Rp-8-Br-PET-cGMP阻断。这些结果表明，通过cGMP介导的PDE 3抑制NO-cAMP相互作用的可能参与。事实上，PDE 3抑制剂西洛酰胺引起的增强8-溴-cAMP-引起的Cx43表达的升高，这是类似于SNAP的效果，并在SNAP处理的细胞中检测到细胞内cAMP的升高。使用基因工程报告MC，表达cAMP反应元件控制下的分泌型碱性磷酸酶，cAMP引起的激活cAMP反应元件的SNAP显着增强。这种效果被废除存在的PDE 3抑制剂西洛酰胺。两者合计，结果表明，NO参与控制GJIC和Cx43的表达。NO的这种作用是由于通过cGMP依赖性抑制PDE 3活性而激活蛋白激酶A。
This study investigated a potential role of nitric oxide (NO) in the regulation of gap junctional intercellular communication (GJIC). Incubation of mesangial cells (MC) with NO donor S-nitroso-N-acetylpenicillamine (SNAP) enhanced both basal and 8-bromo-cAMP-stimulated GJIC as well as expression of gap junction protein connexin43 (Cx43). This potentiating action of SNAP on Cx43 expression was mimicked by two other NO donors and significantly blocked by soluble guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3,-alpha]quinoxalin-1-1. Guanosine 3',5'-cyclic monophosphate (cGMP) analogue 8-bromo-cGMP exerted an effect similar to NO, whereas another cGMP analogue, 8-pCPT-cGMP, which selectively activates GMP-dependent kinase without affecting cGMP-inhibited phosphodiesterase (PDE3), had no effect. Moreover, the synergistic action of NO on Cx43 expression was completely prevented by protein kinase A inhibitor H89 but not by cGMP-dependent kinase inhibitor Rp-8-Br-PET-cGMP. These results suggested a possible involvement of NO-cAMP interaction via cGMP-mediated inhibition of PDE3. Indeed, PDE3 inhibitor cilostamide caused potentiation of 8-bromo-cAMP- elicited elevations of Cx43 expression that is similar to the effect of SNAP, and an elevation of intracellular cAMP was detected in SNAP-treated cells. With the use of genetically engineered reporter MC that express secreted alkaline phosphatase under the control of the cAMP response element, significant potentiation of cAMP-elicited activation of cAMP response element by SNAP was found. This effect was abrogated in the presence of PDE3 inhibitor cilostamide. Taken together, the results suggest that NO is involved in the control of GJIC and Cx43 expression. This effect of NO is due to activation of protein kinase A via cGMP-dependent inhibition of PDE3 activity.