Analysis of the molecular mechanism in regulation of cell migration by alpha2-plasmin inhibitor

纤溶酶原（PG）缺陷小鼠导致细胞迁移受损和伤口愈合延迟。然而，PG缺陷小鼠纤维蛋白原缺乏纠正了愈合时间。因此，这些结果表明，组织纤维蛋白与纤溶蛋白（PM）的降解是细胞迁移的关键。血管抑制素（AST）被认为是一种内源性的新生血管抑制剂。1998年，AST被证明可以抑制内皮细胞的迁移和管状形成。AST是PG的一个片段，含有赖氨酸结合位点（lysine binding site， LBS）， PG和PM通过该位点与纤维蛋白、细胞膜和α 2-纤溶酶抑制剂（alpha2-PI）结合。因此，我们分析了cL2-PI对pm介导的细胞迁移和ast抑制作用的影响。1)α 2- pi参与组织纤维蛋白溶解的机制分析：当纤维蛋白原或纤维蛋白存在于细胞外基质中时，α 2- pi通过Xllla因子与之交联。由于alpha2-PI对选择素有很高的亲和力，我们对alpha2-PI的糖部分进行了更多的分析。cQ-PI经肼降解和唾液酸酶处理后，用gyocomap分析法和阴离子交换高效液相色谱法对其进行了研究。通过RAMM分析和PNA结合推断了低聚糖的结构。结果，alpha2-PI具有含有1-3个唾液酸的n -连接寡糖和含有1个唾液酸的0-连接寡糖。此外，本研究表明α - pl含有一个sialyl- Lewis x结构。2)细胞迁移研究：细胞外基质中含有交联α 2- pi2的抗干扰素可减弱内皮细胞的迁移。此外，α 2- pi延缓了脂多糖刺激内皮细胞的迁移。此外，α 2- pl似乎可以中和AST对细胞迁移的抑制作用，而a-氨基己酸则没有。我们的研究结果表明，成纤维细胞生长因子特异性地与纤维蛋白结合，表明alpha2-PI在伤口愈合的精细调节中起重要作用。少

Plasminogen (PG) deficient mice result in impaired cell migration and display delayed wound healing. However, fibrinogen deficiency in PG deficient mice corrects healing time. Consequently, these results suggest that the degradation of tissue-fibrin with plasmin (PM) is critical for cell migration. Angiostatin (AST) has been identified as an endogenous inhibitor of neovascularization. In 1998, AST has been shown to inhibit migration and tube formation of endothelial cells. AST, a fragment of PG, contains lysine-binding site (LBS) through which PG and PM bind to fibrin, cell membrane and alpha2-plasmin inhibitor (alpha2-PI). Thereby, we have analyzed the effect of cL2-PI on the PM-mediated cell migration and the inhibitory effect of AST.1) Analysis of the mechanism of alpha2-PI to participate in tissue fibrinolysis : When fibrinogen or fibrin existed asan extracellular matrix, alpha2-PI was crosslinked to this by factor Xllla. Since alpha2-PI had a high affinity to selectin, we have ana … More lyzed the sugar moiety of alpha2-PI.After degradation with hydrazine and treatment with sialidase, cQ-PI was studied by gyocomap analysis and anion exchange HPLC.Structure of oligosaccharides was deduced by RAMM analysis and the PNA binding. In result, alpha2-PI has N-linked oligosaccharides which has 1-3 sialic acids and 0-linked one containing one sialic acid. In addition, this study has suggested that alpha-Pl contains a sialyl- Lewis x structure.2) Cell migration studies : The extracellular matrix which contained tibrinogen with crosslinked alpha2-PIattenuated endothelial cell migration. In addition, alpha2-PI delayed migration of endothelial cells, which were stimulated with lipopolysaccharide. Furthermore, alpha2-Pl appeared to neutralize the inhibitory effect of AST on cell migration, whereas a-amino caproic acid did not. Our results with the report that fibroblast growth factor specifically binds to fibrin indicate that alpha2-PI plays an important role in a fine regulation of wound healing. Less