Adenosine A1 receptors promote vasa vasorum endothelial cell barrier integrity via Gi and Akt-dependent actin cytoskeleton remodeling.
Adenosine A1 receptors promote vasa vasorum endothelial cell barrier integrity via Gi and Akt-dependent actin cytoskeleton remodeling.
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DOI：
10.1371/journal.pone.0059733
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Gerasimovskaya EV
中科院分区：
文献类型：
--
作者：
Umapathy SN;Kaczmarek E;Fatteh N;Burns N;Lucas R;Stenmark KR;Verin AD;Gerasimovskaya EV
在缺氧性肺动脉高压的新生儿模型中，观察到显著的肺动脉外膜增厚、炎性细胞在外膜隔室中的积聚以及血管微循环网络的血管生成性扩张。这些病理生理反应表明，快速增殖的血管内皮细胞（VVEC）可能表现出增加的渗透性循环血细胞和大分子。然而，这些观察结果背后的分子机制仍然未被探索。一些报道暗示细胞外腺苷在缺氧和炎症条件下调节血管通透性。因此，我们的目的是确定腺苷的作用，在屏障调节的VVEC分离的肺动脉的常氧（VVEC-Co）或慢性缺氧（VVEC-Hyp）新生小牛。我们通过跨内皮电阻测量表明，外源性腺苷显著增强了VVEC-Co的屏障功能，在较小程度上，在VVEC-Hyp。我们的定量逆转录聚合酶链反应的数据表明，VVEC-Co和VVEC-Hyp表达所有四种腺苷受体（A1，A2 A，A2 B和A3），A1受体（A1 Rs）的表达水平最高。然而，A1 R的表达是显着降低VVEC-Hyp相比，VVEC-Co。通过使用A1 R-特异性激动剂/拮抗剂和siRNA，我们证明，A1 Rs主要负责腺苷诱导的增强屏障功能。用百日咳毒素和GSK 690693或LY 294002预处理VVEC可减弱腺苷诱导的屏障完整性增强，表明Gi蛋白和PI 3 K-Akt通路参与。此外，我们揭示了肌动蛋白细胞骨架在VVEC屏障调节中的关键作用，通过使用肌动蛋白和微管聚合的特异性抑制剂。此外，我们发现腺苷预处理阻断了肿瘤坏死因子α（TNF-α）诱导的VVEC-Co渗透性，验证了其抗炎作用。我们首次证明，A1受体的刺激增强了VVEC的屏障功能，通过激活Gi/PI 3 K/Akt通路和肌动蛋白微丝的重塑。
In a neonatal model of hypoxic pulmonary hypertension, a dramatic pulmonary artery adventitial thickening, accumulation of inflammatory cells in the adventitial compartment, and angiogenic expansion of the vasa vasorum microcirculatory network are observed. These pathophysiological responses suggest that rapidly proliferating vasa vasorum endothelial cells (VVEC) may exhibit increased permeability for circulating blood cells and macromolecules. However, the molecular mechanisms underlying these observations remain unexplored. Some reports implicated extracellular adenosine in the regulation of vascular permeability under hypoxic and inflammatory conditions. Thus, we aimed to determine the role of adenosine in barrier regulation of VVEC isolated from the pulmonary arteries of normoxic (VVEC-Co) or chronically hypoxic (VVEC-Hyp) neonatal calves. We demonstrate via a transendothelial electrical resistance measurement that exogenous adenosine significantly enhanced the barrier function in VVEC-Co and, to a lesser extent, in VVEC-Hyp. Our data from a quantitative reverse transcription polymerase chain reaction show that both VVEC-Co and VVEC-Hyp express all four adenosine receptors (A1, A2A, A2B, and A3), with the highest expression level of A1 receptors (A1Rs). However, A1R expression was significantly lower in VVEC-Hyp compared to VVEC-Co. By using an A1R-specific agonist/antagonist and siRNA, we demonstrate that A1Rs are mostly responsible for adenosine-induced enhancement in barrier function. Adenosine-induced barrier integrity enhancement was attenuated by pretreatment of VVEC with pertussis toxin and GSK690693 or LY294002, suggesting the involvement of Gi proteins and the PI3K-Akt pathway. Moreover, we reveal a critical role of actin cytoskeleton in VVEC barrier regulation by using specific inhibitors of actin and microtubule polymerization. Further, we show that adenosine pretreatment blocked the tumor necrosis factor alpha (TNF-α)-induced permeability in VVEC-Co, validating its anti-inflammatory effects. We demonstrate for the first time that stimulation of A1Rs enhances the barrier function in VVEC by activation of the Gi/PI3K/Akt pathway and remodeling of actin microfilament.
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影响因子：
20.1
作者：
Hasegawa T;Bouïs D;Liao H;Visovatti SH;Pinsky DJ
通讯作者：
Pinsky DJ
影响因子：
20.1
作者：
Kolosova, IA;Mirzapoiazova, T;Verin, AD
通讯作者：
Verin, AD
影响因子：
20.3
作者：
Eltzschig, HK;Thompson, LF;Colgan, SP
通讯作者：
Colgan, SP
影响因子：
158.5
作者：
BARGER, AC;BEEUWKES, R;SILVERMAN, KJ
通讯作者：
SILVERMAN, KJ
DOI：
10.1152/ajplung.00343.2005
发表时间：
2006-08-01
影响因子：
4.9
作者：
Jacobson, Jeffrey R.;Dudek, Steven M.;Garcia, Joe G. N.
通讯作者：
Garcia, Joe G. N.