切应力对与血管平滑肌细胞联合培养的内皮细胞细胞外基质构筑及含量的影响

Changes in Extracellular Matrix of Endothelial Cells Co-cultured with VSMC under Shear Stress

为了探讨切应力对与血管平滑肌细胞联合培养的内皮细胞黏附能力的影响 ,为改进血管内皮细胞种植的组织工程技术提供实验基础 ,我们应用免疫荧光细胞化学、激光共聚焦扫描显微镜和图像分析等技术 ,研究了切应力作用下与血管平滑肌细胞联合培养的内皮细胞的细胞外基质中纤维粘连蛋白、层粘连蛋白和 型胶原的构筑及含量的变化 ,同时以静态条件下联合培养的内皮细胞为对照组。结果表明 ,静态条件下 ,纤维粘连蛋白和层粘连蛋白在细胞内均以颗粒的形式存在于核周区 ,以纤丝的形式存在于细胞外 ,且纤丝相互交织成网状。 型胶原主要以颗粒的形式存在于核周围 ,纤丝极少。切应力作用下纤维粘连蛋白形成纤丝束 ,并有沿切应力方向排列的趋势 ,层粘连蛋白、 型胶原也形成纤丝束 ,但排列无方向性。细胞外基质各组分的含量均有不同程度的变化。本文结果提示 ,切应力作用下 ,与血管平滑肌细胞联合培养的内皮细胞的黏附能力可能增强。

The construction and amount of fibronectin (Fn), laminin (Ln) and collagen type IV (ColIV) in the extracellular matrix of endothelial cells (EC) co cultured with vascular smooth muscle cells (VSMC) under shear stress were studied by immuno fluoro cytochemistry, laser confocal scanning microscopy and image analysis methods to detect the effect of shear stress on adhesion of EC for vascular tissue engineering. One group of EC was maintained under static conditions as a control. In the control, both Fn and Ln exhibited as a granular pattern in perinuclear area and a fibrillar pattern localized underneath EC, and ColⅣ exhibited mainly as the granular in the perinuclear area, but the fibrillar was fewer. On exposure of EC to shear stress in physiological range, Fn grouped into fibril tracts, and there was a tendency for some of these tracts of fibrils to align with direction of shear stress. Ln and ColⅣ also grouped into fibril tracts, which, in contrast to Fn, were randomly oriented. Simultaneously, all contents of the extracellular matrix had different quantitative alterations. These results suggest that the capability of adhesion of EC co cultured with VSMC may be strengthened under shear stress.