Alkaloid rich fraction from Nelumbo nucifera targets VSMC proliferation and migration to suppress restenosis in balloon-injured rat carotid artery
Alkaloid rich fraction from Nelumbo nucifera targets VSMC proliferation and migration to suppress restenosis in balloon-injured rat carotid artery
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DOI：
10.1016/j.atherosclerosis.2016.03.020
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发表时间：
2016-05-01
期刊：
影响因子：
5.3
通讯作者：
Kim, Dong-Wook
中科院分区：
文献类型：
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作者：
Jun, Moon Young;Karki, Rajendra;Kim, Dong-Wook
目的：再狭窄是血管成形术后的不良后果，动脉粥样硬化的特征是血管平滑肌细胞（VSMC）异常增殖和迁移，导致新生内膜形成。本研究观察了莲生物碱富集部位（alkaloid rich fraction，ARF）和复方甲基莲心碱（compound neferine）对血小板源性生长因子（platelet-derived growth factor，PDGF-BB）诱导的VSMC增殖、迁移和大鼠颈动脉损伤模型新生内膜形成的抑制作用。逆转录-聚合酶链反应（RT-PCR）测定细胞周期相关分子的基因表达。Western blot检测细胞内PDGF-R β、细胞外调节激酶（ERK）1/2、c-Jun N末端激酶（JNK）、P38、金属蛋白酶（MMP）-9和核因子-κ B（NF-κ B）等信号分子的表达。使用免疫荧光显微镜评价应力纤维形成。采用大鼠颈动脉球囊损伤模型观察ARF对新生内膜形成的影响。ARF和甲基莲心碱的抗增殖活性是由于抑制细胞周期蛋白D1，细胞周期蛋白E和细胞周期蛋白依赖性激酶（Cdk）基因的表达。此外，ARF和甲基莲心碱抑制PDGF-R β、ERK 1/2、JNK和P38激活以及NF-κ B易位。ARF和甲基莲心碱通过抑制MMP-9活性而抑制VSMC迁移，但不影响细胞骨架重塑。在大鼠颈动脉损伤模型中，ARF抑制新生内膜形成。结论：我们的研究结果表明，ARF通过抑制PDGF-R β介导的信号传导，靶向VSMC增殖和迁移，从而减少新生内膜形成。(C)2016爱思唯尔爱尔兰有限公司版权所有。
Aims: Restenosis-an adverse consequence following angioplasty, and atherosclerosis are characterized by abnormal vascular smooth muscle cell (VSMC) proliferation and migration leading to neo-intima formation. In the present study, we investigated the inhibitory effects of alkaloid rich fraction (ARF) from Nelumbo nucifera and isolated compound neferine on platelet-derived growth factor (PDGF-BB) induced VSMC proliferation and migration in vitro and neo-intima formation in a rat carotid artery injury model.Methods: PDGF-BB induced VSMC proliferation and migration was assessed using colorimetric assay and modified Boyden chamber method respectively. Gene expression of cell cycle associated molecules was determined by reverse transcription-polymerase chain reaction (RT-PCR). The signaling molecules such as PDGF-R beta, extracellular regulated kinase (ERK) 1/2, c-Jun N-terminal kinase (JNK), P38, metalloproteinase (MMP)-9 and nuclear factor-kappa B (NF-kappa B) were determined by western blot analysis. Stress fiber formation was evaluated using immunofluorescence microscopy. The rat carotid artery balloon injury model was performed to assess the effect of ARF on neo-intima formation.Results: ARF possessed the strongest anti-oxidant activities. The anti-proliferative activity of both ARF and neferine was due to suppression of cyclin D1, cyclin E and cyclin-dependent kinase (Cdk) gene expression. Moreover, ARF and neferine inhibited PDGF-R beta, ERK1/2, JNK and P38 activations and NF-kappa B translocation. Also, ARF and neferine inhibited VSMC migration by inhibiting MMP-9 activity without affecting cytoskeleton remodeling. In a rat carotid artery injury model, ARF inhibited neo-intima formation.Conclusion: Our results indicate that ARF targets VSMC proliferation and migration to attenuate neointima formation by inhibition of PDGF-R beta mediated signaling. (C) 2016 Elsevier Ireland Ltd. All rights reserved.