Endothelial cell substrate stiffness influences neutrophil transmigration via myosin light chain kinase-dependent cell contraction
Endothelial cell substrate stiffness influences neutrophil transmigration via myosin light chain kinase-dependent cell contraction
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DOI：
10.1182/blood-2010-11-321125
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发表时间：
2011-08-11
期刊：
影响因子：
20.3
通讯作者：
Aranda-Espinoza, Helim
中科院分区：
文献类型：
--
作者：
Stroka, Kimberly M.;Aranda-Espinoza, Helim
大量的工作已经致力于剪切流和细胞因子对白细胞转运的影响。然而，没有研究探讨基底刚度对迁移的影响。在这里，我们使用体外血管内皮模型研究了内皮细胞收缩介导的中性粒细胞转运的重要方面。我们使用纤维连接蛋白包被的不同生理硬度的聚丙烯酰胺凝胶来模拟不同力学性能的血管，并镀上人脐静脉内皮细胞（HUVEC）单层，这些单层被肿瘤坏死因子- α激活。有趣的是，随着内皮以下底物刚度的增加，中性粒细胞的迁移也增加。HUVEC细胞间粘附分子-1的表达、刚度、细胞骨架排列、形态和细胞-底物粘附不能解释迁移对HUVEC底物刚度的依赖。我们还探讨了细胞收缩的作用，并观察到在中性粒细胞迁移达到最大值几分钟后，在坚硬底物上的内皮细胞形成了大孔。此外，通过抑制肌球蛋白轻链激酶来抑制收缩，通过减少HUVECs在坚硬底物上的迁移和消除孔形成，使底物硬度的影响正常化。这些结果提供了强有力的证据，证明中性粒细胞转运是由肌球蛋白轻链激酶介导的内皮细胞收缩调节的，并且这一事件取决于内皮细胞基质的刚度。(血。2011;118 (6):1632 - 1640)
A vast amount of work has been dedicated to the effects of shear flow and cytokines on leukocyte transmigration. However, no studies have explored the effects of substrate stiffness on transmigration. Here, we investigated important aspects of endothelial cell contraction-mediated neutrophil transmigration using an in vitro model of the vascular endothelium. We modeled blood vessels of varying mechanical properties using fibronectin-coated polyacrylamide gels of varying physiologic stiffness, plated with human umbilical vein endothelial cell (HUVEC) monolayers, which were activated with tumor necrosis factor-alpha. Interestingly, neutrophil transmigration increased with increasing substrate stiffness below the endothelium. HUVEC intercellular adhesion molecule-1 expression, stiffness, cytoskeletal arrangement, morphology, and cell-substrate adhesion could not account for the dependence of transmigration on HUVEC substrate stiffness. We also explored the role of cell contraction and observed that large holes formed in endothelium on stiff substrates several minutes after neutrophil transmigration reached a maximum. Further, suppression of contraction through inhibition of myosin light chain kinase normalized the effects of substrate stiffness by reducing transmigration and eliminating hole formation in HUVECs on stiff substrates. These results provide strong evidence that neutrophil transmigration is regulated by myosin light chain kinase-mediated endothelial cell contraction and that this event depends on subendothelial cell matrix stiffness. (Blood. 2011;118(6):1632-1640)