Synergistic effect of shear stress and streptavidin-biotin on the expression of endothelial vasodilator and cytoskeleton genes.
Synergistic effect of shear stress and streptavidin-biotin on the expression of endothelial vasodilator and cytoskeleton genes.
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DOI：
10.1002/bit.20263
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发表时间：
2004-12
影响因子：
3.8
通讯作者：
B. P. Chan;W. Reichert;G. Truskey
中科院分区：
文献类型：
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作者：
B. P. Chan;W. Reichert;G. Truskey
链霉亲和素（SA）-生物素和纤连蛋白（Fn）的双配体处理增强了内皮细胞（EC）在合成表面上的粘附，并促进粘附EC的静止表型。目前的研究调查的双重配体对内皮基因的表达在静态培养和剪切应力下（4小时，10达因/厘米2）的影响。检测了23个基因在信号传导、细胞骨架/ECM、血管调节和剪切反应类中的表达。8个基因（α-琥珀酸合成酶、K+通道、TGF β、Mn-SOD、α-微管蛋白、t-PA、COX 2和eNOS）被剪切应力显著上调。两个基因（caveolin-1和ET-1）下调切应力。三个基因（RhoA，弹性蛋白，α-辅肌动蛋白）上调的双配体处理在静态培养，和四个基因（FAK，弹性蛋白，COX 2，和eNOS）上调时，双配体和剪切应力同时施加。对FAK、RhoA、弹性蛋白和α-辅肌动蛋白的北方印迹分析显示了类似的结果。结果表明：（1）使用SA-生物素补充EC粘附通过上调细胞骨架/ECM基因的表达增强EC细胞骨架的完整性，以及（2）细胞骨架/ECM基因的表达与下游事件（如剪切诱导的eNOS和COX 2基因的表达）之间可能存在关系。本研究中提出的分析提供了深入了解SA-生物素补充EC介导基因表达的机制。
Dual ligand treatment of streptavidin(SA)-biotin and fibronectin (Fn) enhances the adhesion of endothelial cells (EC) onto synthetic surfaces and promotes the quiescent phenotype of adherent EC. The current study investigates the effect of the dual ligand on the expression of endothelial genes in static culture and under shear stress (4 h at 10 dynes/cm2). Expression of 23 genes in the classes of signaling, cytoskeleton/ECM, vasoregulation, and shear-responsive were examined. Eight genes (argininosuccinate synthetase, K+ channel, TGFbeta, Mn-SOD, alpha-tubulin, t-PA, COX2, and eNOS) were significantly upregulated by shear stress. Two genes (caveolin-1 and ET-1) were downregulated by shear stress. Three genes (RhoA, elastin, alpha-actinin) were upregulated by the dual ligand treatment in static culture, and four genes (FAK, elastin, COX2, and eNOS) were upregulated when the dual ligand and shear stress were applied simultaneously. Northern blot analyses on FAK, RhoA, elastin, and alpha-actinin revealed similar results. The results suggest (1) the use of SA-biotin to supplement EC adhesion enhances the integrity of the EC cytoskeleton by upregulating the expression of cytoskeleton/ECM genes, and (2) a likely relationship between the expression of cytoskeleton/ECM genes and the downstream events, such as the shear-induced expression of eNOS and COX2 genes. Analyses presented in this study provide insights into the mechanism by which SA-biotin-supplemented EC mediate gene expression.