白细胞介素-1 受体相关激酶-1 在大鼠颈动脉损伤后血管平滑肌细胞增殖和新生内膜形成中的作用。
Involvement of interleukin-1 receptor-associated kinase-1 in vascular smooth muscle cell proliferation and neointimal formation after rat carotid injury.
机构信息
From the Pharmacology Division, CSIR-Central Drug Research Institute, Lucknow, India.
出版信息
Arterioscler Thromb Vasc Biol. 2015 Jun;35(6):1445-55. doi: 10.1161/ATVBAHA.114.305028. Epub 2015 Apr 23.
OBJECTIVE
Reduced frequency of atherosclerotic plaques is observed in interleukin-1 receptor-associated kinase-1 (IRAK1)-deficient mice; however, the underlying mechanism is not clear. Therefore, this study investigate the role of IRAK1 in vascular smooth muscle cell proliferation and neointimal hyperplasia.
APPROACH AND RESULTS
Stimulation of rat primary vascular smooth muscle cells with fetal bovine serum (10%) or platelet-derived growth factor-BB (20 ng/mL) for 15 minutes to 24 hours induced a time-dependent increase in IRAK1 and extracellular signal-regulated kinase (ERK) activation, proliferating cell nuclear antigen upregulation and p27Kip1 downregulation as assessed by Western blotting. Inhibitors of ERK pathway (U0126, 10 μmol/L), IRAK (IRAK1/4, 3 μmol/L), protein kinase C (PKC; Ro-31-8220, 1 μmol/L), siRNA of toll-like receptor-4 (200 nmol/L), and PKC-ε (200 nmol/L) significantly attenuated these changes. Platelet-derived growth factor induced endogenous IRAK-ERK-PKC-ε association in a toll-like receptor-4 and PKC-ε-dependent manner. A time-dependent increase in IRAK1 and ERK activation was observed after 15 minutes, 30 minutes, 1 hour, 6 hours, 12 hours, and 24 hours of carotid balloon injury in rats. Balloon injury induced endogenous IRAK-ERK-PKC-ε interaction. Perivascular application of IRAK1/4 inhibitor (100 μmol/L), U0126 (100 μmol/L), and IRAK1 siRNA (220 and 360 nmol/L) in pluronic gel abrogated balloon injury-induced ERK phosphorylation, activation, and p27Kip1 downregulation. Hematoxylin and eosin staining and immunohistochemistry of proliferating cell nuclear antigen and smooth muscle actin demonstrated that balloon injury-induced intimal thickening and neointimal vascular smooth muscle cell proliferation were significantly abrogated in the presence of IRAK1/4 inhibitor, IRAK1 siRNA, and U0126.
CONCLUSIONS
IRAK1 mediates vascular smooth muscle cell proliferation and neointimal hyperplasia by regulating PKC-ε-IRAK1-ERK axis.
目的
在白细胞介素-1 受体相关激酶-1（IRAK1）缺陷小鼠中观察到动脉粥样硬化斑块的频率降低；然而，其潜在机制尚不清楚。因此，本研究旨在探讨 IRAK1 在血管平滑肌细胞增殖和新生内膜增生中的作用。
方法和结果
用胎牛血清（10%）或血小板衍生生长因子-BB（20ng/ml）刺激大鼠原代血管平滑肌细胞 15 分钟至 24 小时，Western 印迹法检测到 IRAK1 和细胞外信号调节激酶（ERK）激活、增殖细胞核抗原上调和 p27Kip1 下调呈时间依赖性增加。ERK 通路抑制剂（U0126，10μmol/L）、IRAK（IRAK1/4，3μmol/L）、蛋白激酶 C（PKC；Ro-31-8220，1μmol/L）、Toll 样受体-4（TLR-4）siRNA（200nmol/L）和 PKC-ε（200nmol/L）显著减弱了这些变化。血小板衍生生长因子诱导内源性 IRAK-ERK-PKC-ε 以 TLR-4 和 PKC-ε 依赖的方式结合。在大鼠颈总动脉球囊损伤后 15 分钟、30 分钟、1 小时、6 小时、12 小时和 24 小时观察到 IRAK1 和 ERK 激活的时间依赖性增加。球囊损伤诱导内源性 IRAK-ERK-PKC-ε 相互作用。在多聚体凝胶中应用 IRAK1/4 抑制剂（100μmol/L）、U0126（100μmol/L）和 IRAK1 siRNA（220 和 360nmol/L）可阻断球囊损伤诱导的 ERK 磷酸化、激活和 p27Kip1 下调。苏木精和伊红染色以及增殖细胞核抗原和平滑肌肌动蛋白的免疫组织化学显示，在 IRAK1/4 抑制剂、IRAK1 siRNA 和 U0126 存在的情况下，球囊损伤诱导的内膜增厚和新生内膜血管平滑肌细胞增殖明显受到抑制。
结论
IRAK1 通过调节 PKC-ε-IRAK1-ERK 轴介导血管平滑肌细胞增殖和新生内膜增生。
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