Kuzuya M, Satake S, Miura H, Hayashi T, Iguchi A
Department of Geriatrics, Nagoya University School of Medicine, Japan.
Exp Cell Res. 1996 Aug 1;226(2):336-45. doi: 10.1006/excr.1996.0234.
The vascular basement membrane is involved in the regulation of endothelial cell differentiation. The accumulation of advanced glycosylation endproducts (AGEs) has been demonstrated on these basement membranes in patients with diabetes. We examined the effect of AGEs on endothelial cell behavior on reconstituted basement membrane, Matrigel. Human umbilical vein-derived endothelial cells (HUVECs) stopped proliferating and differentiated into capillary-like tube-shaped structures on Matrigel. Laminin antibody partially blocked this process. HUVECs cultured on glycosylated Matrigel, however, proliferated and formed a monolayer without tube formation. The inclusion of aminoguanidine, an inhibitor of AGE formation, during the glycosylation of Matrigel restored HUVEC differentiation. Although the laminin adsorbed onto the plastic culture wells promoted HUVEC attachment and spreading, glycosylated laminin reduced HUVEC attachment by 50% and abolished cellular spreading. These effects were restored by aminoguanidine. HUVEC attachment to glycosylated laminin was further reduced by AGE-modified albumin, poly I, acetylated low-density lipoprotein, or maleylated albumin, ligands for a scavenger receptor. Coating the culture dishes with the laminin peptides RGD, YIGSR, and SIKVAV supported the attachment of HUVECs that was unaffected by glycosylation. Results suggest that AGE accumulation on the basement membranes inhibits endothelial cell differentiation by impairing the normal interactions of endothelial cell receptors with their specific matrix ligands. This process may be involved in diabetic angiopathy.
血管基底膜参与内皮细胞分化的调节。在糖尿病患者的这些基底膜上已证实存在晚期糖基化终产物（AGEs）的积累。我们研究了AGEs对内皮细胞在重组基底膜基质胶上行为的影响。人脐静脉来源的内皮细胞（HUVECs）在基质胶上停止增殖并分化为毛细血管样管状结构。层粘连蛋白抗体部分阻断了这一过程。然而，在糖基化的基质胶上培养的HUVECs增殖并形成单层，不形成管状结构。在基质胶糖基化过程中加入氨基胍（一种AGE形成抑制剂）可恢复HUVEC的分化。尽管吸附在塑料培养孔上的层粘连蛋白促进了HUVEC的附着和铺展，但糖基化的层粘连蛋白使HUVEC的附着减少了50%，并消除了细胞铺展。这些作用可被氨基胍恢复。AGE修饰的白蛋白、聚I、乙酰化低密度脂蛋白或马来酰化白蛋白（一种清道夫受体的配体）进一步降低了HUVEC与糖基化层粘连蛋白的附着。用层粘连蛋白肽RGD、YIGSR和SIKVAV包被培养皿可支持HUVEC的附着，且不受糖基化影响。结果表明，基底膜上AGE的积累通过损害内皮细胞受体与其特定基质配体的正常相互作用来抑制内皮细胞分化。这一过程可能与糖尿病血管病变有关。
