Death-associated protein kinase 3 mediates vascular structural remodelling via stimulating smooth muscle cell proliferation and migration.
Death-associated protein kinase 3 mediates vascular structural remodelling via stimulating smooth muscle cell proliferation and migration.
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DOI：
10.1042/cs20130591
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发表时间：
2014-10
期刊：
影响因子：
6
通讯作者：
T. Usui;Tomoki Sakatsume;Ryo Nijima;Kosuke Otani;K. Kazama;T. Morita;S. Kameshima;M. Okada;H. Yamawaki
中科院分区：
文献类型：
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作者：
T. Usui;Tomoki Sakatsume;Ryo Nijima;Kosuke Otani;K. Kazama;T. Morita;S. Kameshima;M. Okada;H. Yamawaki
死亡相关蛋白激酶 3 (DAPK3) 也称为拉链相互作用激酶，是一种丝氨酸/苏氨酸激酶，主要调节细胞死亡和平滑肌收缩。我们之前发现自发性高血压大鼠（SHR）肠系膜动脉中 DAPK3 的蛋白表达增加，并且 DAPK3 部分通过促进活性氧依赖性血管炎症来介导 SHR 高血压的发生。然而，DAPK3如何控制平滑肌细胞（SMC）增殖和迁移仍有待阐明，这也是高血压发展的重要过程。因此，我们试图研究 DAPK3 是否影响 SMC 增殖和迁移。分别通过溴脱氧尿苷 (BrdU) 掺入和细胞计数测定以及 Boyden 室测定确定，针对 DAPK3 的 siRNA 显着抑制血小板衍生生长因子 (PDGF)-BB 诱导的 SMC 增殖和迁移。通过罗丹明-鬼笔环肽染色测定，DAPK3 siRNA 或 DAPK3 的药理学抑制剂抑制 PDGF-BB 诱导的片状伪足形成。通过蛋白质印迹法测定，DAPK3 siRNA 或 DAPK 抑制剂显着降低 PDGF-BB 诱导的 p38 和热休克蛋白 27 (HSP27) 的激活。在离体研究中，PDGF-BB 诱导的 SMC 生长被 DAPK 抑制剂显着抑制。在体内，DAPK 抑制剂在小鼠结扎模型中显着预防颈动脉新生内膜增生。目前的结果首次揭示DAPK3通过激活p38/HSP27信号介导PDGF-BB诱导的SMC增殖和迁移，这可能导致血管结构重塑，包括新生内膜增生。本研究表明 DAPK3 作为预防高血压心血管疾病的新药物靶点。
Death-associated protein kinase 3 (DAPK3) also known as zipper-interacting kinase is a serine/threonine kinase that mainly regulates cell death and smooth muscle contraction. We have previously found that protein expression of DAPK3 increases in the mesenteric artery from spontaneously hypertensive rats (SHRs) and that DAPK3 mediates the development of hypertension in SHRs partly through promoting reactive oxygen species-dependent vascular inflammation. However, it remains to be clarified how DAPK3 controls smooth muscle cell (SMC) proliferation and migration, which are also important processes for hypertension development. We, therefore, sought to investigate whether DAPK3 affects SMC proliferation and migration. siRNA against DAPK3 significantly inhibited platelet-derived growth factor (PDGF)-BB-induced SMC proliferation and migration as determined by bromodeoxyuridine (BrdU) incorporation and a cell counting assay as well as a Boyden chamber assay respectively. DAPK3 siRNA or a pharmacological inhibitor of DAPK3 inhibited PDGF-BB-induced lamellipodia formation as determined by rhodamine-phalloidin staining. DAPK3 siRNA or the DAPK inhibitor significantly reduced PDGF-BB-induced activation of p38 and heat-shock protein 27 (HSP27) as determined by Western blotting. In ex vivo studies, PDGF-BB-induced SMC out-growth was significantly inhibited by the DAPK inhibitor. In vivo, the DAPK inhibitor significantly prevented carotid neointimal hyperplasia in a mouse ligation model. The present results, for the first time, revealed that DAPK3 mediates PDGF-BB-induced SMC proliferation and migration through activation of p38/HSP27 signals, which may lead to vascular structural remodelling including neointimal hyperplasia. The present study suggests DAPK3 as a novel pharmaceutical target for the prevention of hypertensive cardiovascular diseases.