Extracellular beta-nicotinamide adenine dinucleotide (beta-NAD) promotes the endothelial cell barrier integrity via PKA- and EPAC1/Rac1-dependent actin cytoskeleton rearrangement.
Extracellular beta-nicotinamide adenine dinucleotide (beta-NAD) promotes the endothelial cell barrier integrity via PKA- and EPAC1/Rac1-dependent actin cytoskeleton rearrangement.
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DOI：
10.1002/jcp.22029
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发表时间：
2010-04
影响因子：
5.6
通讯作者：
Verin, Alexander D.
中科院分区：
文献类型：
--
作者：
Umapathy, Nagavedi S.;Zemskov, Evgeny A.;Gonzales, Joyce;Gorshkov, Boris A.;Sridhar, Supriya;Chakraborty, Trinad;Lucas, Rudolf;Verin, Alexander D.
已知细胞外β-NAD升高钙离子、肌醇1，4，5-三磷酸和cAMP的细胞内水平。最近，β-NAD被鉴定为P2 Y1和P2 Y11嘌呤能受体的激动剂。由于β-NAD可以从内皮细胞（EC）细胞外释放，我们认为它参与了EC通透性的调节。在此，我们第一次发现，内皮完整性在内皮细胞内源性表达β-NAD激活的嘌呤能受体时，β-NAD刺激可以增强。我们的数据表明，细胞外β-NAD以浓度依赖性方式增加人肺动脉EC（HPAEC）单层的跨内皮电阻（TER），表明内皮屏障增强。重要的是，β-NAD显著减弱凝血酶诱导的EC通透性以及革兰氏阴性和革兰氏阳性细菌毒素的屏障损害作用，这代表β-NAD的屏障保护功能。免疫荧光显微镜显示细胞-细胞连接蛋白VE-钙粘蛋白在细胞外周的染色更明显，表明β-NAD刺激后细胞-细胞接触的紧密性增加。有趣的是，抑制分析（药理学拮抗剂和受体序列特异性siRNA）表明P2 Y1和P2 Y11受体均参与β-NAD诱导的TER增加。B-NAD处理减弱脂多糖（LPS）诱导的肌球蛋白轻链磷酸化，表明其参与屏障保护。我们的研究还表明cAMP依赖性蛋白激酶A和EPAC 1途径以及小GTdR Rac 1参与β-NAD诱导的EC屏障增强。根据这些结果，我们得出结论，β-NAD通过小的GTdR Rac 1和MLCP依赖性信号通路调节肺EC屏障完整性。
Extracellular β-NAD is known to elevate intracellular levels of calcium ions, inositol 1,4,5-trisphate and cAMP. Recently, β-NAD was identified as an agonist for P2Y1 and P2Y11 purinergic receptors. Since β-NAD can be released extracellularly from endothelial cells (EC), we have proposed its involvement in the regulation of EC permeability. Here we show, for the first time, that endothelial integrity can be enhanced in EC endogenously expressing β-NAD-activated purinergic receptors upon β-NAD stimulation. Our data demonstrate that extracellular β-NAD increases the transendothelial electrical resistance (TER) of human pulmonary artery EC (HPAEC) monolayers in a concentration-dependent manner indicating endothelial barrier enhancement. Importantly, β-NAD significantly attenuated thrombin-induced EC permeability as well as the barrier-compromising effects of Gram-negative and Gram-positive bacterial toxins representing the barrier-protective function of β-NAD. Immunofluorescence microscopy reveals more pronounced staining of cell-cell junctional protein VE-cadherin at the cellular periphery signifying increased tightness of the cell-cell contacts after β-NAD stimulation. Interestingly, inhibitory analysis (pharmacological antagonists and receptor sequence specific siRNAs) indicates the participation of both P2Y1 and P2Y11 receptors in β-NAD-induced TER increase. B-NAD-treatment attenuates the lipopolysaccharide (LPS)-induced phosphorylation of myosin light chain indicating its involvement in barrier protection. Our studies also show the involvement of cAMP-dependent protein kinase A and EPAC1 pathways as well as small GTPase Rac1 in β-NAD-induced EC barrier enhancement. With these results, we conclude that β-NAD regulates the pulmonary EC barrier integrity via small GTPase Rac1- and MLCP- dependent signaling pathways.
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影响因子：
4.8
作者：
Dudek, S. M.;Camp, S. M.;Singleton, P. A.
通讯作者：
Singleton, P. A.
影响因子：
2.1
作者：
Lum, H;Malik, AB
通讯作者：
Malik, AB
影响因子：
5.3
作者：
Fukuhara, S;Sakurai, A;Mochizuki, N
通讯作者：
Mochizuki, N
影响因子：
20.1
作者：
Kolosova, IA;Mirzapoiazova, T;Verin, AD
通讯作者：
Verin, AD
DOI：
10.1152/ajpheart.00542.2005
发表时间：
2006-01-01
影响因子：
4.8
作者：
Baffert, F;Le, T;McDonald, DM
通讯作者：
McDonald, DM