Molecular mechanisms mediating protective effect of cAMP on lipopolysaccharide (LPS)-induced human lung microvascular endothelial cells (HLMVEC) hyperpermeability.
Molecular mechanisms mediating protective effect of cAMP on lipopolysaccharide (LPS)-induced human lung microvascular endothelial cells (HLMVEC) hyperpermeability.
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DOI：
10.1002/jcp.21913
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发表时间：
2009-12
影响因子：
5.6
通讯作者：
Verin, Alexander D.
中科院分区：
文献类型：
--
作者：
Bogatcheva, Natalia V.;Zemskova, Marina A.;Kovalenkov, Yevgeniy;Poirier, Christophe;Verin, Alexander D.
迄今为止，脓毒症引起的血管渗漏的性质仅被部分理解，这限制了其管理的药理学方法。在这里，我们研究了环磷酸腺苷的保护作用，使用内毒素诱导的高通透性作为模型的屏障功能障碍，观察革兰氏阴性脓毒症。我们证明，减轻LPS诱导的屏障损害可以通过特异性激活蛋白激酶A（PKA）或Epac与cAMP类似物Bnz-cAMP或O-Me-cAMP，分别实现。我们接下来研究了PKA底物VASP和filamin 1在屏障维持和LPS诱导的屏障损害中的参与。VASP和filamin 1与特定的siRNA的消耗显着加剧了静止细胞屏障和LPS诱导的屏障功能障碍，这表明这些蛋白质的屏障保护作用。VASP耗竭与更严重的损失ZO-1外周染色响应LPS，而filamin 1耗竭细胞反应LPS与更强大的应力纤维诱导和更深刻的变化ZO-1和VE-钙粘蛋白外周组织。VASP和filamin 1磷酸化均显著增加，这是PKA激活的结果。我们接下来分析了VASP和细丝蛋白1消耗对LPS诱导的屏障损害的PKA依赖性缓解的影响。我们观察到，Bnz-cAMP的能力，以抵消LPS诱导的高通透性衰减只有VASP，但没有filamin 1耗尽。我们的数据表明，虽然PKA依赖的VASP磷酸化有助于cAMP引起的LPS受损的单层细胞的保护作用，filamin 1磷酸化不太可能在这个过程中发挥重要作用。
Up to date, the nature of the sepsis-induced vascular leakage is understood only partially, which limits pharmacological approaches for its management. Here we studied the protective effect of cAMP using endotoxin-induced hyperpermeability as a model for barrier dysfunction observed in gram-negative sepsis. We demonstrated that the alleviation of LPS-induced barrier compromise could be achieved by the specific activation of either protein kinase A (PKA) or Epac with cAMP analogues Bnz-cAMP or O-Me-cAMP, respectively. We next studied the involvement of PKA substrates VASP and filamin1 in barrier maintenance and LPS-induced barrier compromise. Depletion of both VASP and filamin1 with the specific siRNAs significantly exacerbated both the quiescent cells barrier and LPS-induced barrier dysfunction, suggesting barrier-protective role of these proteins. VASP depletion was associated with the more severe loss of ZO-1 peripheral staining in response to LPS, whereas filamin1-depleted cells reacted to LPS with more robust stress fiber induction and more profound changes in ZO-1 and VE-cadherin peripheral organization. Both VASP and filamin1 phosphorylation was significantly increased as a result of PKA activation. We next analyzed the effect of VASP and filamin1 depletion on the PKA-dependent alleviation of LPS-induced barrier compromise. We observed that Bnz-cAMP ability to counteract LPS-induced hyperpermeability was attenuated only by VASP, but not filamin1 depletion. Our data indicate that while PKA-dependent VASP phosphorylation contributes to the protective effect of cAMP elicited on LPS-compromised monolayers, filamin1 phosphorylation is unlikely to play a significant role in this process.
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DOI：
10.1158/1078-0432.ccr-08-1402
发表时间：
2009-02-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Bedolla RG;Wang Y;Asuncion A;Chamie K;Siddiqui S;Mudryj MM;Prihoda TJ;Siddiqui J;Chinnaiyan AM;Mehra R;de Vere White RW;Ghosh PM
通讯作者：
Ghosh PM
DOI：
10.1152/ajplung.00417.2007
发表时间：
2008-04-01
影响因子：
4.9
作者：
Rentsendorj, Otgonchimeg;Mirzapoiazova, Tamara;Pearse, David B.
通讯作者：
Pearse, David B.
影响因子：
5.5
作者：
Schlegel, N.;Burger, S.;Waschke, J.
通讯作者：
Waschke, J.
影响因子：
20.1
作者：
Kolosova, IA;Mirzapoiazova, T;Verin, AD
通讯作者：
Verin, AD
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.