Functional analysis of LR11 in vascular smooth muscle cells

LR11在血管平滑肌细胞中的功能分析

• 批准号: 12835002
• 负责人: BUJO Hideaki
• 金额: $ 2.37万
• 依托单位: CHIBA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12835002/
• 关键词: LDLR; LR11; Atherosclerosis; Smooth muscle cells; 動脈硬化; LDL受容体; 血管

LR11, a member of the LDL receptor family, is highly expressed in vascular smooth muscle cells (SMCs) of the hyperplastic intima, but not media. To further clarify the involvement of LR11 in the process of atherosclerosis, we have characterized the migration and invasion activities, of LR11-overexpressing SMCs. LR11 cDNA was transfected into the rat SMC line, A7r5. Compared to mock cells (C-1), in the presence of PDGF-BB the transfected cells (R-1 and R-2) showed 3.5-4.0-fold higher expression of LR11 protein, 1.7-1.8-fold increased migration, and 2.0-2.2-fold elevated invasion activities, respectively. The increases were essentially abolished by the addition of receptor-associated protein (RAP), anti-LR11 antibodies, or Apo E. Immunological analyzes showed that urokinase-type plasminogen activator receptor (uPAR) levels were increased in LR11-overexpressing cells. Anti-uPA and anti-uPAR antibodies reduced the migration and invasion activities of R-1 and R-2 cells to baseline levels. RAP, anti-ER11 antibodies, and apo E decreased uPAR expression in the LR11-overexpressing cells by 〜50%. Cellular catabolism of uPAR was significantly decreased in R-1 and R-2 cells compared to control. Cultured SMCs isolated from intimae of atherosclerotic rabbit aortas showed increased expression levels of LR11 and uPAR, and enhanced migration and invasion compared to SMCs from medial layers. Overexpression of LR11 induces enhanced migration and invasion activities of intimal SMCs in vitro, likely via its regulation of the uPA/uPAR system.

LR11 是 LDL 受体家族的成员，在增生性内膜的血管平滑肌细胞 (SMC) 中高表达，但在中膜中不表达。为了进一步阐明LR11在动脉粥样硬化过程中的参与，我们对LR11过表达的SMCs的迁移和侵袭活性进行了表征。 LR11 cDNA 转染至大鼠 SMC 系 A7r5 中。与模拟细胞（C-1）相比，在存在PDGF-BB的情况下，转染细胞（R-1和R-2）的LR11蛋白表达分别提高了3.5-4.0倍，迁移能力提高了1.7-1.8倍，侵袭活性提高了2.0-2.2倍。通过添加受体相关蛋白 (RAP)、抗 LR11 抗体或 Apo E，这种增加基本上被消除。免疫学分析表明，在 LR11 过表达细胞中，尿激酶型纤溶酶原激活剂受体 (uPAR) 水平增加。抗uPA和抗uPAR抗体将R-1和R-2细胞的迁移和侵袭活性降低至基线水平。 RAP、抗 ER11 抗体和 apo E 使 LR11 过表达细胞中 uPAR 表达降低约 50%。与对照相比，R-1 和 R-2 细胞中 uPAR 的细胞分解代谢显着降低。与来自内层的 SMC 相比，从动脉粥样硬化兔主动脉内膜分离培养的 SMC 表现出 LR11 和 uPAR 的表达水平增加，并且迁移和侵袭增强。 LR11 的过度表达可能通过调节 uPA/uPAR 系统，在体外诱导内膜 SMC 的迁移和侵袭活性增强。

项目成果

Miyazaki O, Kobayashi J, Fukamachi I, Miida T, Bujo H, Saito Y.: "A new sandwich enzyme immunoassay for measurement of plasma pre-betal-HDL level"J Lipid Res.. 41(12). 2083-2088 (2000)

Miyazaki O、Kobayashi J、Fukamachi I、Miida T、Bujo H、Saito Y.：“一种用于测量血浆前 β-HDL 水平的新型夹心酶免疫测定法”J Lipid Res.. 41(12)。

Kanaki T, Morisaki N, Bujo H, Takahashi K, Ishii I, Saito Y: "The regulatory expression of procollagen COOH-terminal proteinase enhancer in the proliferation of vascular smooth muscle cells."Biochem Biophys Res Commun. 270(3). 1049-54 (2000)

Kanaki T、Morisaki N、Bujo H、Takahashi K、Ishii I、Saito Y：“前胶原 COOH 末端蛋白酶增强剂在血管平滑肌细胞增殖中的调节表达。”Biochem Biophys Res Commun。

Hirayama S, Bujo H, Yamazaki H, Kanaki T, Takahashi K, Saito Y.: "Differential expression of LR11 during proliferation and differentiation of cultured ~neuroblastoma cells"Biochem Biophys Res Commun. 275(2). 365-562 (2000)

Hirayama S、Bujo H、Yamazaki H、Kanaki T、Takahashi K、Saito Y.：“培养的神经母细胞瘤细胞增殖和分化过程中 LR11 的差异表达”Biochem Biophys Res Commun。

Zhu Y., et al.: "Enhanced expression of LDLR family member LR11 increases migration of smooth muscle cells in vitro"Circulation. In press. (2002)

Zhu Y.等人：“LDLR家族成员LR11的表达增强可增加平滑肌细胞的体外迁移”循环。

Tanaga K, et al.: "Incteased circulating MDA-LDL levels in patients with coronary artery diseases and its association with the peak sizes of LDL particles"Arterioscler.Thromb.Vasc.Biol. In press. (2002)

Tanaga K 等人：“冠状动脉疾病患者循环 MDA-LDL 水平的增加及其与 LD​​L 颗粒峰值大小的关系”Arterioscler.Thromb.Vasc.Biol。