Patterns of living beta-actin movement in wounded human coronary artery endothelial cells exposed to shear stress.
Patterns of living beta-actin movement in wounded human coronary artery endothelial cells exposed to shear stress.
复制标题
暴露于剪切应力的受伤人冠状动脉内皮细胞中活β-肌动蛋白的运动模式。
DOI：
10.1006/excr.2001.5351
复制
发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Flozak,AS
中科院分区：
文献类型：
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作者：
Albuquerque,ML;Flozak,AS
我们之前证明了生理水平的剪切应力可以增强内皮细胞的修复。细胞的扩散和迁移，而不是增殖，是伤口愈合率增加的主要机制(Albuquerque et al., 2000, Am。j .杂志。中华心脏杂志，2003,19(3):393 - 393。然而，在人冠状动脉内皮细胞（HCAECs）中，负责细胞运动和易位的β-肌动蛋白丝的模式和运动在生理流动下尚未被研究过。转染β-actin-GFP的hcaec在I型胶原包被盖层上培养。融合细胞单层在平行板流室中承受12达因/平方厘米的层流剪切应力18小时，以达到细胞排列，然后用金属刮刀刮伤，随后暴露在20达因/平方厘米的层流剪切应力（S-W-sH）或静态（S-W-sT）条件下。在施加S-W-sH或S-W-sT条件后的前3小时进行延时成像和反褶积显微镜检查。分析了两种条件下HCAEC单层膜伤口愈合过程中β-actin-GFP运动和易位的时空动态。与S-W-sT条件下的HCAEC相比，我们的数据显示，S-W-sH条件下的HCAEC表现出更大的β-actin-GFP运动性，纤维和团块模式，以及细胞附着和脱离过程中使用的纤维弧。这些发现证明了β-肌动蛋白在暴露于生理血流的HCAEC伤口愈合过程中的组织和运动的有趣模式。
We previously demonstrated that physiologic levels of shear stress enhance endothelial repair. Cell spreading and migration, but not proliferation, were the major mechanisms accounting for the increases in wound closure rate (Albuquerque et al., 2000, Am. J. Physiol. Heart Circ. Physiol. 279, H293–H302). However, the patterns and movements of β-actin filaments responsible for cell motility and translocation in human coronary artery endothelial cells (HCAECs) have not been previously investigated under physiologic flow. HCAECs transfected with β-actin-GFP were cultured on type I collagen-coated coverslips. Confluent cell monolayers were subjected to laminar shear stress of 12 dynes/cm2for 18 h in a parallel-plate flow chamber to attain cellular alignment and then wounded by scraping with a metal spatula and subsequently exposed to a laminar shear stress of 20 dynes/cm2(S-W-sH) or static (S-W-sT) conditions. Time-lapse imaging and deconvolution microscopy was performed during the first 3 h after imposition of S-W-sH or S-W-sT conditions. The spatial and temporal dynamics of β-actin-GFP motility and translocation during wound closure in HCAEC monolayers were analyzed under both conditions. Compared with HCAEC under S-W-sT conditions, our data show that HCAEC under S-W-sH conditions demonstrated greater β-actin-GFP motility, filament and clumping patterns, and filament arcs used during cellular attachment and detachment. These findings demonstrate intriguing patterns of β-actin organization and movement during wound closure in HCAEC exposed to physiological flow.