Keegan Philip M, Anbazhakan Suhaas, Kang Baolin, Pace Betty S, Platt Manu O
Biol Chem. 2016 May;397(5):459-68. doi: 10.1515/hsz-2015-0244.
Cathepsins K and V are powerful elastases elevated in endothelial cells by tumor necrosis factor-α (TNFα) stimulation and disturbed blood flow both of which contribute to inflammation-mediated arterial remodeling. However, mechanisms behind endothelial cell integration of biochemical and biomechanical cues to regulate cathepsin production are not known. To distinguish these mechanisms, human aortic endothelial cells (HAECs) were stimulated with TNFα and exposed to pro-remodeling or vasoprotective shear stress profiles. TNFα upregulated cathepsin K via JNK/c-jun activation, but vasoprotective shear stress inhibited TNFα-stimulated cathepsin K expression. JNK/c-jun were still phosphorylated, but cathepsin K mRNA levels were significantly reduced to almost null indicating separate biomechanical regulation of cathepsin K by shear stress separate from biochemical stimulation. Treatment with Bay 11-7082, an inhibitor of IκBα phosphorylation, was sufficient to block induction of cathepsin K by both pro-remodeling shear stress and TNFα, implicating NF-κB as the biomechanical regulator, and its protein levels were reduced in HAECs by vasoprotective shear stress. In conclusion, NF-κB and AP-1 activation were necessary to activate cathepsin K expression in endothelial cells, highlighting integration of biochemical and biomechanical stimuli to control cathepsins K and V, powerful elastases implicated for arterial remodeling due to chronic inflammation and disturbed blood flow.
组织蛋白酶K和V是强大的弹性蛋白酶,在肿瘤坏死因子-α(TNFα)刺激和血流紊乱的情况下,内皮细胞中它们的水平会升高,这两者都有助于炎症介导的动脉重塑。然而,内皮细胞整合生化和生物力学信号以调节组织蛋白酶产生的机制尚不清楚。为了区分这些机制,用人主动脉内皮细胞(HAECs)进行实验,用TNFα刺激并使其暴露于促重塑或血管保护性剪切应力模式下。TNFα通过JNK/c-jun激活上调组织蛋白酶K,但血管保护性剪切应力抑制TNFα刺激的组织蛋白酶K表达。JNK/c-jun仍然被磷酸化,但组织蛋白酶K的mRNA水平显著降低至几乎为零,这表明剪切应力对组织蛋白酶K有独立于生化刺激的生物力学调节作用。用IκBα磷酸化抑制剂Bay 11-7082处理足以阻断促重塑剪切应力和TNFα对组织蛋白酶K的诱导,这表明NF-κB是生物力学调节因子,并且血管保护性剪切应力会降低HAECs中其蛋白水平。总之,NF-κB和AP-1的激活对于激活内皮细胞中组织蛋白酶K的表达是必要的,这突出了生化和生物力学刺激的整合对组织蛋白酶K和V的控制作用,这两种强大的弹性蛋白酶与慢性炎症和血流紊乱导致的动脉重塑有关。