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

α2-纤溶酶抑制剂调控细胞迁移的分子机制分析

• 批准号: 09470234
• 负责人: SAKATA Yoichi
• 金额: $ 3.01万
• 依托单位: Jichi Medical School, School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470234/
• 关键词: plasminogen; alpha2-plasmin inhibitor; angiostatin; fibrin; factor XIII; selectin; α2プラスミンインヒビタ; 血管内皮細胞; 架橋結合; プラスミノゲン; 細胞移動; 組織修復

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

纤溶酶原（PG）缺陷小鼠导致细胞迁移受损并显示伤口愈合延迟。然而，PG缺陷小鼠中的纤维蛋白原缺陷纠正了愈合时间。因此，这些结果表明，纤溶酶（PM）的组织纤维蛋白的降解是细胞迁移的关键。血管抑素（Angiostatin，AST）是一种内源性的新生血管抑制剂。1998年，AST已被证明可抑制内皮细胞的迁移和管形成。AST是PG的一个片段，含有赖氨酸结合位点（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（α 2-PI）结合。1）α_2-PI参与组织纤溶的机制分析：当纤维蛋白原或纤维蛋白以细胞外基质形式存在时，α_2-PI通过因子XIIIa与之交联。由于α 2-PI对选择素有很高的亲和力，我们用α 2-PI对选择素进行了分析。 ...更多信息 将α_2-PI的糖部分裂解，用肼和唾液酸酶降解后，用Gyocomap分析和阴离子交换HPLC分析，用RAMM分析和PNA结合来推测寡糖的结构。因此，α 2-PI具有含1-3个唾液酸的N-连接寡糖和含1个唾液酸的O-连接寡糖。此外，本研究还表明α-PI含有唾液酸-刘易斯X结构。2）细胞迁移研究：含有与交联的α 2-PI的骨蛋白原的细胞外基质减弱内皮细胞迁移。此外，α 2-PI延迟迁移的内皮细胞，这是刺激脂多糖。此外，α 2-Pl似乎中和AST对细胞迁移的抑制作用，而α-氨基己酸则没有。我们的研究结果与报告，成纤维细胞生长因子特异性结合纤维蛋白表明，α 2-PI在伤口愈合的精细调节中起着重要的作用。少

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