Seo Hyang-Hee, Kim Sang Woo, Lee Chang Youn, Lim Kyu Hee, Lee Jiyun, Lim Soyeon, Lee Seahyoung, Hwang Ki-Chul
Brain Korea 21 PLUS Project for Medical Science, Yonsei University, Seoul, Korea.
Institute for Bio-Medical Convergence, College of Medicine, Catholic Kwandong University, Gangneung-si, Gangwon-do, Korea.
Eur J Pharmacol. 2017 Mar 5;798:35-42. doi: 10.1016/j.ejphar.2017.02.004. Epub 2017 Feb 6.
Excessive vascular smooth muscle cell (VSMC) proliferation and migration after vascular injury significantly contributes to the development of occlusive vascular disease. Therefore, inhibiting the proliferation and migration of VSMCs is a validated therapeutic modality for occlusive vascular disease such as atherosclerosis and restenosis. In the present study, we screened chemical compounds for their anti-proliferative effects on VSMCs using multiple approaches, such as MTT assays, wound healing assays, and trans-well migration assays. Our data indicate that 7-cyclopentyl-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d] pyrimidin-4-ylamine, a lymphocyte-specific protein tyrosine kinase (Lck) inhibitor, significantly inhibited both VSMC proliferation and migration. 7-cyclopentyl-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine suppresses VSMC proliferation and migration via down-regulating the protein kinase B (Akt) and extracellular signal regulated kinase (ERK) pathways, and it significantly decreased the expression of proliferating cell nuclear antigen (PCNA) and cyclin D1 and, the phosphorylation of retinoblastoma protein (pRb). Additionally, 7-cyclopentyl-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d] pyrimidin-4-ylamine suppressed the migration of VSMCs from endothelium-removed aortic rings, as well as neointima formation following rat carotid balloon injury. The present study identified 7-cyclopentyl-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine as a potent VSMC proliferation and migration inhibitor and warrants further studies to elucidate its more detailed molecular mechanisms, such as its primary target, and to further validate its in vivo efficacy as a therapeutic agent for pathologic vascular conditions, such as restenosis and atherosclerosis.
血管损伤后血管平滑肌细胞（VSMC）过度增殖和迁移是导致闭塞性血管疾病发展的重要因素。因此，抑制VSMC的增殖和迁移是治疗诸如动脉粥样硬化和再狭窄等闭塞性血管疾病的有效治疗方式。在本研究中，我们采用多种方法，如MTT试验、伤口愈合试验和Transwell迁移试验，筛选对VSMC具有抗增殖作用的化合物。我们的数据表明，淋巴细胞特异性蛋白酪氨酸激酶（Lck）抑制剂7-环戊基-5-（4-苯氧基苯基）-7H-吡咯并[2,3-d]嘧啶-4-胺能显著抑制VSMC的增殖和迁移。7-环戊基-5-（4-苯氧基苯基）-7H-吡咯并[2,3-d]嘧啶-4-胺通过下调蛋白激酶B（Akt）和细胞外信号调节激酶（ERK）信号通路来抑制VSMC的增殖和迁移，并且它能显著降低增殖细胞核抗原（PCNA）和细胞周期蛋白D1的表达以及视网膜母细胞瘤蛋白（pRb）的磷酸化水平。此外，7-环戊基-5-（4-苯氧基苯基）-7H-吡咯并[2,3-d]嘧啶-4-胺抑制了VSMC从去内皮主动脉环的迁移以及大鼠颈动脉球囊损伤后的新生内膜形成。本研究确定7-环戊基-5-（4-苯氧基苯基）-7H-吡咯并[2,3-d]嘧啶-4-胺是一种有效的VSMC增殖和迁移抑制剂，值得进一步研究以阐明其更详细的分子机制，如主要靶点，并进一步验证其作为治疗再狭窄和动脉粥样硬化等病理性血管疾病的体内疗效。
