Shear-induced endothelial NOS activation and remodeling via heparan sulfate, glypican-1, and syndecan-1.
Shear-induced endothelial NOS activation and remodeling via heparan sulfate, glypican-1, and syndecan-1.
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DOI：
10.1039/c3ib40199e
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发表时间：
2014-03
期刊：
影响因子：
--
通讯作者：
Tarbell JM
中科院分区：
文献类型：
--
作者：
Ebong EE;Lopez-Quintero SV;Rizzo V;Spray DC;Tarbell JM
哺乳动物上皮细胞被多功能表面糖萼（GCX）包被。在血管内皮细胞（EC）上，完整的GCX具有动脉粥样硬化保护作用。它在许多血管疾病中降解。GCX硫酸乙酰肝素（HS）对于健康的血流诱导的EC一氧化氮（NO）释放、延伸和对齐至关重要。HS核心蛋白参与这些过程的机制是未知的。我们假设磷脂酰肌醇蛋白聚糖-1（GPC1）HS核心蛋白介导流动诱导的EC NO合酶（eNOS）激活，因为GPC1锚定到eNOS所在的小窝。我们还推测HS核心蛋白syndecan-1（SDC1）介导流动诱导的EC伸长和排列，因为SDC1与影响细胞形状的细胞骨架相连。我们通过暴露用HS降解肝素酶III（HepIII）处理的EC单层和用RNA沉默的GPC1或SDC1处理的EC单层至3至24小时的生理剪切应力来测试我们的假设。剪切条件下的EC与完整的GCX表现出特征性的eNOS激活在短期流动条件。在长期暴露后，具有完整GCX的EC被拉长并沿流动方向排列。HS去除和GPC1抑制，而不是SDC1减少，阻断剪切诱导的eNOS激活。EC重塑响应流量衰减HS降解和SDC1的情况下，但保留与GPC1敲低。这些研究结果清楚地表明，HS参与集中和分散GCX介导的机械传导机制，GPC1作为一个集中的机械传导剂和SDC1在分散的机械传导功能。这项基础性工作展示了EC如何将流体剪切力转化为不同的生物分子和生物力学响应。
Mammalian epithelial cells are coated with a multifunctional surface glycocalyx (GCX). On vascular endothelial cells (EC), intact GCX is atheroprotective. It is degraded in many vascular diseases. GCX heparan sulfate (HS) is essential for healthy flow-induced EC nitric oxide (NO) release, elongation, and alignment. The HS core protein mechanisms involved in these processes are unknown. We hypothesized that the glypican-1 (GPC1) HS core protein mediates flow-induced EC NO synthase (eNOS) activation because GPC1 is anchored to caveolae where eNOS resides. We also hyphothesized that the HS core protein syndecan-1 (SDC1) mediates flow-induced EC elongation and alignment because SDC1 is linked to the cytoskeleton which impacts cell shape. We tested our hypotheses by exposing EC monolayers treated with HS degrading heparinase III (HepIII), and monolayers with RNA-silenced GPC1, or SDC1, to 3 to 24 hours of physiological shear stress. Shear-conditioned EC with intact GCX exhibited characteristic eNOS activation in short-term flow conditions. After long-term exposure, EC with intact GCX were elongated and aligned in the direction of flow. HS removal and GPC1 inhibition, not SDC1 reduction, blocked shear-induced eNOS activation. EC remodeling in response to flow was attenuated by HS degradation and in the absence of SDC1, but preserved with GPC1 knockdown. These findings clearly demonstrate that HS is involved in both centralized and decentralized GCX-mediated mechanotransduction mechanisms, with GPC1 acting as a centralized mechanotransmission agent and SDC1 functioning in decentralized mechanotransmission. This foundational work demonstrates how EC can transform fluid shear forces into diverse biomolecular and biomechanical responses.
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DOI：
10.1016/j.bbrc.2007.01.137
发表时间：
2007-03-30
影响因子：
3.1
作者：
Pahakis, Manolis Y.;Kosky, Jason R.;Tarbell, John M.
通讯作者：
Tarbell, John M.
DOI：
10.1083/jcb.200404171
发表时间：
2004-10-11
期刊：
The Journal of cell biology
影响因子：
--
作者：
Beauvais DM;Burbach BJ;Rapraeger AC
通讯作者：
Rapraeger AC
影响因子：
4
作者：
McQuade, Kyle J.;Beauvais, DeannaLee M.;Rapraeger, Alan C.
通讯作者：
Rapraeger, Alan C.
DOI：
10.1073/pnas.93.13.6448
发表时间：
1996-06-25
影响因子：
11.1
作者：
GarciaCardena, G;Oh, P;Sessa, WC
通讯作者：
Sessa, WC
影响因子：
5.5
作者：
Koo, Andrew;Dewey, C. Forbes, Jr.;Garcia-Cardena, Guillermo
通讯作者：
Garcia-Cardena, Guillermo