WCU Biomodulation Major, Department of Agricultural Biotechnology and Center for Food and Bioconvergence, Seoul National University, Seoul 151-921, Republic of Korea.
Cardiovasc Res. 2014 Mar 1;101(3):503-12. doi: 10.1093/cvr/cvt337. Epub 2013 Dec 20.
Abnormal migration of human aortic smooth muscle cells (HASMCs) causes intimal thickening of the aorta, a pivotal step in atherosclerotic development. Although many studies have demonstrated that high anthocynidins intake confers protective effects against atherosclerosis, the direct molecular targets, and mechanisms of action responsible remain unclear. Here, we investigated the preventive effect of anthocyanidins on atherosclerosis and the underlying mechanisms involved.
We analysed six major anthocyanidins, and found that petunidin exhibited the most potent inhibitory effects against platelet-derived growth factor (PDGF)-BB-induced HASMC migration in the Boyden chamber and wound healing assays. Petunidin also suppressed PDGF-BB-induced ex vivo rat aortic sprouting and in vivo rat neointima formation. Western blot analysis showed that petunidin inhibited PDGF-BB-induced phosphorylation of focal adhesion kinase (FAK) at the low concentration of 5 µM, whereas phosphorylation of Src, mitogen-activated protein kinases, and Akt was only slightly inhibited at 20 µM. In vitro and ex vivo FAK activity assays demonstrated that petunidin directly suppresses FAK activity by binding in an ATP non-competitive manner. Moreover, anthocyanidins that reduced HASMC migration also inhibited PDGF-BB-induced FAK phosphorylation, F-actin reduction, and FAK activity, and directly bound with FAK. PDGF-BB-induced migration, F-actin reduction by HASMCs, and ex vivo aortic sprouting were all inhibited by treatment with a commercial FAK inhibitor, PF-228.
The results of the present study demonstrate that anthocyanidins can directly bind with and suppress the activity of FAK with atherosclerosis-preventive effects.
人类主动脉平滑肌细胞（HASMCs）的异常迁移导致主动脉内膜增厚，这是动脉粥样硬化发展的关键步骤。尽管许多研究表明，高花青素摄入量对动脉粥样硬化具有保护作用，但负责的直接分子靶点和作用机制仍不清楚。在这里，我们研究了花青素对动脉粥样硬化的预防作用及其涉及的潜在机制。
我们分析了六种主要的花青素，发现矢车菊素对血小板衍生生长因子（PDGF）-BB 诱导的 HASMC 在 Boyden 室和划痕愈合实验中的迁移具有最强的抑制作用。矢车菊素还抑制了 PDGF-BB 诱导的大鼠离体主动脉芽生和大鼠体内新生内膜形成。Western blot 分析表明，矢车菊素在 5µM 的低浓度下抑制 PDGF-BB 诱导的黏着斑激酶（FAK）磷酸化，而在 20µM 时仅略微抑制Src、丝裂原活化蛋白激酶和 Akt 的磷酸化。体外和离体 FAK 活性测定表明，矢车菊素通过非 ATP 竞争方式结合来直接抑制 FAK 活性。此外，降低 HASMC 迁移的花青素也抑制了 PDGF-BB 诱导的 FAK 磷酸化、F-肌动蛋白减少和 FAK 活性，并与 FAK 直接结合。FAK 抑制剂 PF-228 处理可抑制 PDGF-BB 诱导的 HASMC 迁移、F-肌动蛋白减少和离体主动脉芽生。
本研究结果表明，花青素可以直接与 FAK 结合并抑制其活性，具有预防动脉粥样硬化的作用。
