Analysis of the mechanism of angiogenesis regulated by fibrinolytic factors

纤溶因子调控血管生成的机制分析

• 批准号: 13670049
• 负责人: MATSUO Osamu
• 金额: $ 2.24万
• 依托单位: Kinki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670049/
• 关键词: fibrinolysis; u-PA; t-PA; plasminogen; knockout mouse; angiogenesis; gene expression; MMP-2; 線溶系因子; 創傷治癒モデル; カプセル; フィブリン; 線溶系; 線溶系因子欠損マウス; 血管内皮細胞; マトリックスメタロプロテアーゼ

Double gene deficient mouse of the fibrinolytic factors, which lost two kinds of fibrinolytic genes, was produced by crossing single gene deficient mouse of fibrinolytic factor which we have already established. Double gene deficient mice produced were mouse which lost u-PA and u-PAR genes (u-PA/u-PAR double knockout mouse), that lost u-PA and t-PA genes (u-PA/t-PA double knockout mouse), that lost α2-AP and u-PA genes (α2-AP/u-PA double knockout mouse) and th at lost α2-AP and t-PA genes (α2-A P/t-PA double knocktout mouse). The present study was performed in accordance with the institutional guidelines.When the hemispherical capsule, which had many holes, was transplanted to subcutaneous of u-PA/t-PA double knockout mouse, the MMP-2 activity of the granulation tissue, which adhered out side of capsule, was significantly lower than the wild-type mouse. On the other hand, the MMP-2 activity of the granulation tissue, which proliferated in the capsule, was significantly higher than the wild-type mouse.The vascular endothelial cell in granulation tissue which adhered to the capsule in u-PA/t-PA double knockout mouse was fewer than the wild type mouse, and it was indicated that both u-PA and t-PA were involved in the angiogenesis. Since plasminogen is the substrate for u-PA and t-PA, it is suggested that the plasminogen is important for the angiogenesis. Therefore, regulation of expression of the plasminogen gene was examined using the primary cultured liver cell. In the low-density culture, the expression of plasminongen mRNA was decreased. However, in the high-density culure where one hepatocyte attached to another, decrease in plasminogen mRNA was not observed. Therefore, it is confirmed that the stimulation by cell-cell contact may play an important role to maintain the expression of plasminongen mRNA in hepatocytes.

纤溶因子双基因缺陷小鼠是通过与已建立的纤溶因子单基因缺陷小鼠杂交，产生的两种纤溶基因缺失的纤溶因子双基因缺陷小鼠。双基因缺失小鼠包括u-PA和u-PAR基因缺失小鼠（u-PA/u-PAR双基因敲除小鼠）、u-PA和t-PA基因缺失小鼠（u-PA/t-PA双基因敲除小鼠）、α2-AP和u-PA基因缺失小鼠（α2-AP/u-PA双基因敲除小鼠）和α2-AP和t-PA基因缺失小鼠（α2-AP/t-PA双基因敲除小鼠）。本研究按照机构指南进行，当将具有许多孔的半球形囊移植到u-PA/t-PA双基因敲除小鼠的皮下时，粘附在囊外侧的肉芽组织的MMP-2活性显著低于野生型小鼠。另一方面，在被膜内增生的肉芽组织中MMP-2活性明显高于野生型小鼠，而在u-PA/t-PA双基因敲除小鼠中，与被膜粘附的肉芽组织中血管内皮细胞数量明显少于野生型小鼠，表明u-PA和t-PA均参与了血管生成。由于纤溶酶原是u-PA和t-PA的底物，提示纤溶酶原在血管生成中起重要作用。因此，使用原代培养的肝细胞检查纤溶酶原基因表达的调节。在低密度培养中，纤溶酶原mRNA表达减少。而在高密度培养中，两个肝细胞相互贴壁，纤溶酶原mRNA的表达未见减少。因此，证实了细胞-细胞接触刺激可能在维持肝细胞纤溶酶原mRNA表达中起重要作用。

项目成果

Matsuno H., Kozawa O., Yoshimi N., Akamatsu S., Hara A., Mori H., Okada K., Ueshima S., Matsuo O., Uematsu T.: "Lack of α2-antiplasmin promotes pulmonary heart failure via over-release of VEGF after acute myocardial infarction"Blood. 100(7). 2487-2493 (20

Matsuno H.、Kozawa O.、Yoshimi N.、Akamatsu S.、Hara A.、Mori H.、Okada K.、Ueshima S.、Matsuo O.、Uematsu T.：“缺乏 α2-抗纤溶酶会促进肺心力衰竭通过急性心肌梗死后 VEGF 的过度释放“Blood. 100(7). 2487-2493 (20

Matsuno H.: "Lack of α2-antiplasmin promotes pulmonary heart failure via over-release of VEGF after acute myocardial infarction"Blood. 100・7. 2487-2493 (2002)

Matsuno H.：“急性心肌梗死后缺乏 α2-抗纤溶酶通过过度释放 VEGF 促进肺心力衰竭”100・7 2487-2493 (2002)。

Luttun A.: "Loss of placental growth factor protects mice against vascular permeability in pathological conditions"Biochem Blophys Res Commun. 295・2. 428-434 (2002)

Luttun A.：“胎盘生长因子的丧失可保护小鼠免受病理条件下的血管通透性”Biochem Blophys Res Commun. 295·2 428-434 (2002)。

Akao M., Ueshima S., Okada K., Fukao H., Seki T., Ariga T., Matsuo O.: "Cellular density regulation of plasminogen gene expression in mouse hepatocytes"Life Sciences. (in press).

Akao M.、Ueshima S.、Okada K.、Fukao H.、Seki T.、Ariga T.、Matsuo O.：“小鼠肝细胞中纤溶酶原基因表达的细胞密度调节”生命科学。

Ueshima S., Matsuno H., Hayashi M., Horibuchi K., Okada K., Fukao H., Uematsu T., Matsuo O.: "Function of tissue-type plasminogen activator releaser on vascular endothelial cells and thrombolysis in vivo"Thromb Haemostasis. 87(6). 1069-1074 (2002)

Ueshima S.、Matsuno H.、Hayashi M.、Horibuchi K.、Okada K.、Fukao H.、Uematsu T.、Matsuo O.：“组织型纤溶酶原激活物释放剂对血管内皮细胞和体内溶栓的作用”