Calpain-1 Activity and Central Arterial Aging

Calpain-1 活性和中枢动脉老化

• 批准号: 7732167
• 负责人: Edward Lakatta
• 金额: $ 21.26万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7732167
• 关键词: Adenoviruses; Age; Aging; Angiotensin II; Angiotensin II Signaling Pathway; Animal Model; Atherosclerosis; Blood Vessels; Calpain; Cardiac Myocytes; Cells; Collagen; Collagen Type I; Complementary DNA; Cytoskeleton; Disease; Ectopic Expression; Elderly; Endopeptidases; Event; Extracellular Matrix; Fibroblasts; Gelatinase A; Genes; Human; Hypertension; Immigration; In Vitro; Inflammation; Lead; Length; Link; MMP2 gene; Matrix Metalloproteinases; Mediating; Molecular; Osteonectin; Peptide Hydrolases; Phenotype; Play; Production; Proteins; Recombinants; Risk Factors; Role; Signal Transduction; Smooth Muscle Myocytes; Stroke; Transcript; aged; arterial remodeling; calcification; calpain inhibitor; in vivo; osteopontin; prevent; therapy design; vascular smooth muscle cell migration

Angiotensin II (Ang II) signaling, including matrix metalloproteinase type II (MMP2) activation, has been linked to an age-associated increase in migration capacity of vascular smooth muscle cells (VSMC), and to other proinflammatory features of arterial aging. Calpain-1 activation is required for MMP2 expression in fibroblasts and is induced in cardiomyocytes by Ang II. The consequences of engagement of calpain-1 with its substrates, however, in governing the age-associated proinflammatory and procalcification status within the arterial wall, remain unknown. In this study, our results first demonstrate an age-associated increase of calpain-1 gene transcripts, protein levels and activity in the old aortic wall; increased Ang II induces calpain-1 expression in the aortic walls in vivo and ex vivo and VSMC in vitro; the Ang II mediated age-associated increased MMP2 activity, is blocked by calpain inhibitor 1 or CAST, indicating that calpain activity is required for MMP activation by Ang II treatment. An increase in old VSMC migratory capacity is mimicked in young VSMC by over-expression of calpain-1. Collectively, these findings point to increased calpain-1 activity, as a central component of the exaggerated Ang II signaling pathway, which is involved in age-associated arterial remodeling. Thus, Calpain-1 activation is a pivotal molecular event in the age-associated arterial Ang II/MMP2 signaling cascade that is linked to cytoskeleton protein restructuring, and VSMC migration. Furthermore, age-associated arterial alterations are characterized both by a shift of VSMC from a contractile to synthetic phenotype, i.e., cytoskeleton restructuring and enhancement of migratory capability, and extracellular matrix remodeling (ECM), ie., collagen types I and III, which could be degenerated by proteases and become immature collagen to form a procalcification niche. Our recent study indicates that calpain-1, increases within VSMC; and orchestrates an angiotensin II cascade which mediates a VSMC phenotype shift within the aged aortic wall, and plays an important role in generating arterial matrix alterations that lead to procalcification. Our results show that over-expression of ectopic calpain-1 by a recombinant adenovirus harboring full length cDNA (pAd/CANP1) in young VSMC increased Col I and III up to the levels of untreated old control cells. Further, over-expression of calpain-1 in young VSMC decreases the counter-calcification molecules osteopontin (OPN) and osteonectin (ON), which mimics aging. In contrast, young VSMC infected with recombinant adenovirus containing calpastatain cDNA (pAd/CAST), an endogenous inhibitor of calpain), increases OPN and ON. An age-associated increase of calpain-1 activity within VSMCs results in enhanced collagen production and reduced counter-calcification molecules, favoring a procacification status, which is a potential molecular mechanism of arterial matrix remodeling with aging. Thus, targeting calpain-1 is a potential approach to delay or reverse the age-associated arterial pro-inflammation and -calcification state that is also associated with diseases such as hypertension, atherosclerosis, and stroke.

血管紧张素II（ANG II）信号传导，包括基质金属蛋白酶II型（MMP2）激活，与年龄相关的血管平滑肌细胞（VSMC）的迁移能力增加与其他动脉衰老的促炎特征有关。 Calpain-1激活是成纤维细胞中MMP2表达所必需的，并且由ANG II在心肌细胞中诱导。然而，Calpain-1及其底物的参与后果在管理动脉壁内与年龄相关的促炎和分化状态的后果尚不清楚。 在这项研究中，我们的结果首先证明了旧主动脉壁中Calpain-1基因转录物，蛋白质水平和活性的年龄相关。 ANG II的增加会在体内诱导主动脉壁的Calpain-1表达，并在体外和Ex Vivo和VSMC体外诱导Calpain-1。 ANG II介导的与年龄相关的MMP2活性增加，被钙蛋白酶抑制剂1或铸造阻塞，表明通过ANG II治疗激活MMP需要钙蛋白酶活性。通过过度表达CalPain-1，在Young VSMC中模仿了旧VSMC迁移能力的增加。总的来说，这些发现指出了增加Calpain-1活性，这是夸张的ANG II信号通路的中心成分，该途径与年龄相关的动脉重塑有关。因此，Calpain-1激活是与年龄相关的动脉ANG II/MMP2信号级联反应的关键分子事件，与细胞骨架蛋白重组和VSMC迁移有关。 Furthermore, age-associated arterial alterations are characterized both by a shift of VSMC from a contractile to synthetic phenotype, i.e., cytoskeleton restructuring and enhancement of migratory capability, and extracellular matrix remodeling (ECM), ie., collagen types I and III, which could be degenerated by proteases and become immature collagen to form a procalcification利基。我们最近的研究表明，Calpain-1在VSMC内增加。并策划了血管紧张素II级联反应，该级联体介导了老年主动脉壁内的VSMC表型转移，并在产生导致推测的动脉基质改变中起着重要作用。我们的结果表明，在Young VSMC中携带全长cDNA（PAD/CANP1）的重组腺病毒对异位Calpain-1的过表达增加了Col I和III至未经处理的旧对照细胞的水平。 此外，在YOUNG VSMC中的Calpain-1过表达可降低模拟衰老的反估计分子骨桥蛋白（OPN）和骨蛋白切除素（OPN）和骨蛋白切除素（ON）。相比之下，感染了含有Calpastatain cDNA（PAD/CAST）的重组腺病毒（一种Calpain的内源性抑制剂）的年轻VSMC，增加了OPN和ON。 VSMC中Calpain-1活性与年龄相关的增加会导致胶原蛋白的产生增强和降低的反估计分子，有利于推出状态，这是动脉基质重塑随老化的潜在分子机制。因此，靶向CalPain-1是一种潜在的方法，可以延迟或扭转与年龄相关的动脉促炎和 - 定位状态，它也与高血压，动脉粥样硬化和中风等疾病有关。

项目成果

Increased aortic calpain-1 activity mediates age-associated angiotensin II signaling of vascular smooth muscle cells.

• DOI: 10.1371/journal.pone.0002231
• 发表时间: 2008-05-21
• 期刊: PLOS ONE
• 影响因子: 3.7
• 作者: Jiang, Liqun;Wang, Mingyi;Zhang, Jing;Monticone, Robert E.;Telljohann, Richard;Spinetti, Gaia;Pintus, Gianfranco;Lakatta, Edward G.
• 通讯作者: Lakatta, Edward G.