Sun R J, Muller S, Wang X, Zhuang F Y, Stoltz J F
Angiohematology-Hemorheology, LEMTA-UMR-CNRS 7563, Faculté de Médecine, Vandoeuvre-lès-Nancy, France.
Clin Hemorheol Microcirc. 2000;23(1):1-11.
The effect of laminar flow on the regulation of von Willebrand Factor (vWF) of cultured human umbilical vein endothelial cells (HUVECs) was studied. Confluent endothelial monolayers were exposed to shear stresses (0.2 and 1.0 Pa) from 2 to 24 h. vWF was labelled with indirect immunofluorescence method and observed with 3D fluorescence microscopy. The distribution of vWF and the cytoskeleton organization were observed simultaneously by double fluorescence labelling. More actin stress fibers and an increased release of vWF appeared in the cells exposed to flow at the same time. The qualitative and quantitative results showed that there was not only a shear-dependent regulation but also a time-dependent modification. For a short-time shear stimulation, both 0.2 Pa and 1.0 Pa shear stresses induced a release of vWF from the endothelial cells. In contrast, after 24 h exposure to 1.0 Pa shear flow, vWFs were much more in the cells than that in the cells exposed to 0.2 Pa for 24 h (p < 0.01) or that in the control cells (p < 0.05). TNF-alpha caused a decrease of vWF and Weibel-Palade bodies in the cells.
研究了层流对培养的人脐静脉内皮细胞（HUVECs）中血管性血友病因子（vWF）调节的影响。将汇合的内皮细胞单层暴露于2至24小时的剪切应力（0.2和1.0 Pa）下。采用间接免疫荧光法标记vWF，并用三维荧光显微镜观察。通过双重荧光标记同时观察vWF的分布和细胞骨架组织。同时，暴露于流动的细胞中出现了更多的肌动蛋白应力纤维和vWF释放增加。定性和定量结果表明，不仅存在剪切依赖性调节，还存在时间依赖性修饰。对于短时间的剪切刺激，0.2 Pa和1.0 Pa的剪切应力均诱导内皮细胞释放vWF。相反，在暴露于1.0 Pa剪切流24小时后，细胞中的vWF比暴露于0.2 Pa 24小时的细胞（p < 0.01）或对照细胞（p < 0.05）中的vWF多得多。肿瘤坏死因子-α导致细胞中vWF和魏尔-帕拉德小体减少。
