Evidence that NO/cGMP/PKG signalling cascade mediates endothelium dependent inhibition of IP₃R mediated Ca²⁺ oscillations in myocytes and pericytes of ureteric microvascular network in situ.
Evidence that NO/cGMP/PKG signalling cascade mediates endothelium dependent inhibition of IP₃R mediated Ca²⁺ oscillations in myocytes and pericytes of ureteric microvascular network in situ.
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DOI：
10.1016/j.ceca.2015.08.006
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发表时间：
2015-12
期刊：
影响因子：
4
通讯作者：
Burdyga T
中科院分区：
文献类型：
--
作者：
Borysova L;Burdyga T
ODQ（SGC）的抑制剂ODQ逆转了肌细胞和周细胞中Ca2+振荡的内皮依赖性抑制。
选择性PKG抑制剂RP-8-PCPT-CGMP，反向肌细胞和周细胞中激动剂诱导的Ca2+振荡的内皮依赖性终止。
选择性PKG激活剂8PCPT-CGMP诱导激动剂诱导的肌细胞和周细胞中的Ca2+振荡抑制。
Zaprinast显着增强了SNAP的抑制作用。
NO/CGMP/PKG级联反应的抑制作用与抑制Ca2+通过肌细胞和周细胞的IP3R释放有关。
内皮中的Ca2+信号传导与小动脉和静脉周围的肌细胞和周细胞中Ca2+振荡之间的拮抗关系涉及一氧化氮（NO），但潜在的机制涉及一氧化氮。在本研究中，我们研究了Carbachol和没有供体SNAP对使用共聚焦显微镜在完整的肠道肠道微血管网络中的CA2+信号传导和血管舒缩反应的影响。 AVP诱导的动脉和静脉的血管舒张症反应与肌细胞和周细胞中Ca2+振荡的发生相关，并且不会因从细胞外液中去除Ca2+而废除。 Carbachol诱导的内皮细胞内Ca2+的升高伴随着肌细胞和周细胞中Ca2+振荡的终止。这种卡巴醇诱导的抑制作用对肌细胞和周细胞中Ca2+振荡的抑制作用被ODQ（可溶性鸟叶兰氏酶（SGC）的抑制剂）逆转，RP-8-PCPT-CGMPs和蛋白质激酶G（PKG）的抑制剂RP-8-PCPT-CGMP。没有供体捕捉的肌细胞和周细胞中的Ca2+振荡也有效地阻塞。 Zaprinast是CGMP特异性磷酸二酯酶5的选择性抑制剂Zaprinast显着增强了SNAP的抑制作用，并由SGC抑制剂ODQ和PKG抑制剂RP-8-PCPT-CGMP逆转。 CGMP模拟和选择性PKG激活剂8pcpt-CGMP还诱导了肌细胞和周细胞中AVP诱导的Ca2+振荡的抑制。 SNAP对咖啡因在分布街机小动脉中诱导的Ca2+振荡没有影响。因此，我们得出的结论是，在肌细胞和周细胞中，无介导的Ca2+振荡抑制作用主要募集CGMP/PKG依赖性途径。 NO/CGMP/PKG级联反应的抑制作用与通过三磷酸三磷酸盐受体（IP3R）通道选择性地从肌细胞和周细胞中抑制Ca2+释放有关。
Endothelium-dependent inhibition of Ca2+ oscillations in myocytes and pericytes was reversed by ODQ, an inhibitor of soluble guanylyl cyclase (sGC). Selective PKG inhibitor Rp-8-pCPT-cGMPS, reversed endothelium- dependent termination of agonist-induced Ca2+ oscillations in myocytes and pericytes. Selective PKG activator 8pCPT-cGMP induced inhibition of the agonist-induced Ca2+ oscillations in myocytes and pericytes. Inhibitory effect of SNAP was markedly enhanced by zaprinast. Inhibitory effect of NO/cGMP/PKG cascade is associated with suppressed Ca2+ release via IP3Rs of myocytes and pericytes. In ureteric microvessels the antagonistic relationship between Ca2+ signalling in endothelium and Ca2+ oscillations in myocytes and pericytes of arterioles and venules involves nitric oxide (NO), but the underlying mechanisms are not well understood. In the present study we investigated the effects of carbachol and NO donor SNAP on Ca2+ signalling and vasomotor responses of arterioles and venules in intact urteric microvascular network in situ using confocal microscopy. Vasomotor responses of arterioles and venules induced by AVP correlated with the occurrence of Ca2+ oscillations in the myocytes and pericytes and were not abolished by the removal of Ca2+ from extracellular fluid. Carbachol-induced rise of intracellular Ca2+ in endothelium was accompanied by the termination of the Ca2+ oscillations in myocytes and pericytes. This carbachol-induced inhibitory effect on Ca2+ oscillations in myocytes and pericytes was reversed by ODQ, an inhibitor of soluble guanylyl cyclase (sGC) and by Rp-8-pCPT-cGMPS, an inhibitor of protein kinase G (PKG). Ca2+ oscillations in myocytes and pericytes were also effectively blocked by NO donor SNAP. An Inhibitory effect of SNAP was markedly enhanced by zaprinast, a selective inhibitor of cGMP-specific phosphodiesterase-5, and reversed by sGC inhibitor, ODQ and PKG inhibitor, Rp-8-pCPT-cGMPS. The cGMP analogue and selective PKG activator 8pCPT-cGMP also induced inhibition of the AVP-induced Ca2+ oscillations in myocytes and pericytes. SNAP had no effects on Ca2+ oscillations induced by caffeine in distributing arcade arterioles. Consequently, we conclude that NO- mediated inhibition of Ca2+ oscillations in myocytes and pericytes predominantly recruits the cGMP/PKG dependent pathway. The inhibitory effect of NO/cGMP/PKG cascade is associated with suppressed Ca2+ release from the SR of myocytes and pericytes selectively via the inositol triphosphate receptor (IP3R) channels.