The response of fenestrations, actin, and caveolin-1 to vascular endothelial growth factor in SK Hep1 cells
The response of fenestrations, actin, and caveolin-1 to vascular endothelial growth factor in SK Hep1 cells
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DOI：
10.1152/ajpgi.00069.2008
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发表时间：
2008-07-01
影响因子：
4.5
通讯作者：
Le Couteur, David G.
中科院分区：
文献类型：
--
作者：
Cogger, Victoria C.;Arias, Irwin M.;Le Couteur, David G.
为研究血管内皮生长因子对肝窦内皮细胞开窗的调控作用，将绿色荧光蛋白（GFP）-肌动蛋白和GFP-caveolin-1分别转染SK Hep 1细胞。SK Hep 1细胞有气孔;其中一些似乎是穿孔（直径55 ± 28 nm，孔隙率2.0 ± 1.4%）、未发育的筛板、刷毛覆盖的微胞饮囊泡，并表达小窝蛋白-1、血管性血友病因子、血管内皮生长因子受体-2、内皮一氧化氮合酶和网格蛋白，但不表达CD 31。甲醛血清白蛋白被大量摄取，与内吞作用一致。血管内皮生长因子导致孔隙率增加至4.8 ± 2.6%（P < 0.01），孔径增加至104 ± 59 nm（P < 0.001）。GFP-肌动蛋白在整个细胞中表达，而GFP-小窝蛋白1具有点状外观;两者都对血管内皮生长因子通过在数小时内与开窗形成平行的向细胞核收缩作出反应。SK Hep 1细胞类似于肝窦内皮细胞，并且血管内皮生长因子诱导的开窗样孔的形成之前，肌动蛋白细胞骨架和附着的小窝蛋白-1朝向细胞核收缩。
To study the regulation of fenestrations by vascular endothelial growth factor in liver sinusoidal endothelial cells, SK Hep1 cells were transfected with green fluorescence protein (GFP)-actin and GFP-caveolin-1. SK Hep1 cells had pores; some of which appeared to be fenestrations ( diameter 55 +/- 28 nm, porosity 2.0 +/- 1.4%), rudimentary sieve plates, bristlecoated micropinocytotic vesicles and expressed caveolin-1, von Willebrand factor, vascular endothelial growth factor receptor-2, endothelial nitric oxide synthase and clathrin, but not CD31. There was avid uptake of formaldehyde serum albumin, consistent with endocytosis. Vascular endothelial growth factor caused an increase in porosity to 4.8 +/- 2.6% ( P < 0.01) and pore diameter to 104 +/- 59 nm ( P < 0.001). GFP-actin was expressed throughout the cells, whereas GFP-caveolin1 had a punctate appearance; both responded to vascular endothelial growth factor by contraction toward the nucleus over hours in parallel with the formation of fenestrations. SK Hep1 cells resemble liver sinusoidal endothelial cells, and the vascular endothelial growth factor-induced formation of fenestration-like pores is preceded by contraction of actin cytoskeleton and attached caveolin-1 toward the nucleus.