Chen C S, Hawiger J
Department of Medicine, New England Deaconess Hospital, Boston, MA.
Blood. 1991 May 15;77(10):2200-6.
Vascular endothelial cells, providing a nonthrombogenic surface to the lumenal aspect of blood vessels, are anchored to matrix adhesion molecules in the subendothelium through their respective receptors belonging to a superfamily of integrins. We analyzed the reactivity of synthetic peptide analogs of adhesive proteins toward human umbilical vein endothelial cells (HUVEC), assaying their detachment from extracellular matrix and attachment to extracellular matrix components in vitro. Synthetic peptide analogs Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP), Arg-Gly-Asp-Val (RGDV), Arg-Gly-Asp-Ser (RGDS), and Arg-Gly-Asp-Phe (RGDF), which are analogous to "cell adhesion sites" of fibronectin, vitronectin, von Willebrand factor, and alpha-chain of human fibrinogen, respectively, caused significant detachment of HUVEC from the extracellular matrix in vitro at the concentrations ranging from 0.5 to 1.5 mmol/L. They also interfered with attachment of HUVEC to surfaces coated with subendothelial extracellular matrix or its components. The synthetic peptide analog of HHLGGAKQAGDV, which is homologous to the gamma-chain of human fibrinogen sequence 400-411, did not cause any measurable effect on the integrity of HUVEC monolayers (detachment and attachment). "Hybrid" peptides bearing salient features of both sequences, ie, Ala-Lys-Gln-Arg-Gly-Asp-Phe (AKQRGDF) and Lys-Gln-Arg-Gly-Asp-Phe (KQRGDF), had an attenuated effect on the detachment of HUVEC from extracellular matrix. Thus, the integrity of the human endothelial cell monolayer anchored to the extracellular matrix, as measured in detachment and attachment assays, is disturbed by peptides containing RGD sequence whereas the synthetic peptide His-His-Leu-Gly-Gly-Ala-Lys-Gln-Ala-Gly-Asp-Val (HHLGGAKQAGDV) is nonreactive.
血管内皮细胞为血管腔面提供了一个非血栓形成表面，通过属于整合素超家族的各自受体锚定在基底膜下的基质黏附分子上。我们分析了黏附蛋白的合成肽类似物对人脐静脉内皮细胞（HUVEC）的反应性，在体外测定它们从细胞外基质的脱离以及对细胞外基质成分的附着情况。合成肽类似物甘氨酸 - 精氨酸 - 甘氨酸 - 天冬氨酸 - 丝氨酸 - 脯氨酸（GRGDSP）、精氨酸 - 甘氨酸 - 天冬氨酸 - 缬氨酸（RGDV）、精氨酸 - 甘氨酸 - 天冬氨酸 - 丝氨酸（RGDS）和精氨酸 - 甘氨酸 - 天冬氨酸 - 苯丙氨酸（RGDF），它们分别类似于纤连蛋白、玻连蛋白、血管性血友病因子和人纤维蛋白原α链的“细胞黏附位点”，在0.5至1.5 mmol/L的浓度范围内，可在体外导致HUVEC从细胞外基质显著脱离。它们还干扰了HUVEC对包被有基底膜下细胞外基质或其成分的表面的附着。与人纤维蛋白原序列400 - 411的γ链同源的HHLGGAKQAGDV合成肽类似物，对HUVEC单层的完整性（脱离和附着）没有产生任何可测量的影响。具有两个序列显著特征的“杂合”肽，即丙氨酸 - 赖氨酸 - 谷氨酰胺 - 精氨酸 - 甘氨酸 - 天冬氨酸 - 苯丙氨酸（AKQRGDF）和赖氨酸 - 谷氨酰胺 - 精氨酸 - 甘氨酸 - 天冬氨酸 - 苯丙氨酸（KQRGDF），对HUVEC从细胞外基质的脱离作用减弱。因此，在脱离和附着试验中所测量的，锚定在细胞外基质上的人内皮细胞单层的完整性，会受到含RGD序列的肽的干扰，而合成肽组氨酸 - 组氨酸 - 亮氨酸 - 甘氨酸 - 甘氨酸 - 丙氨酸 - 赖氨酸 - 谷氨酰胺 - 丙氨酸 - 甘氨酸 - 天冬氨酸 - 缬氨酸（HHLGGAKQAGDV）无反应。
