Carelli-Alinovi Cristiana, Pirolli Davide, Giardina Bruno, Misiti Francesco
Biochemistry and Clinical Biochemistry Institute, Catholic University, School of Medicine, Rome, Italy.
Istituto di Chimica del Riconoscimento Molecolare (ICRM), National Research Council (CNR), Rome, Italy.
Clin Hemorheol Microcirc. 2015;59(4):345-54. doi: 10.3233/CH-141845.
We have previously showed that morphological alterations in Red Blood Cells (RBCs) are correlated to an impaired eNOS enzymatic activity and a concomitant reduced NO derived metabolites formation. Here we extend our previous observations, reporting that RBC morphology is regulated by a series of specific cell signaling events linked to Protein Kinase C (PKC)-mediated activation of caspase 3. Pretreatment of RBCs with the PKC inhibitor chelerythrine, prior to the addition of phorbol-12-myristate-13-acetate (PMA), an activator of PKC, blocks the appearance of the morphology alterations and the sustained decrease in nitrates and nitrites levels induced by PMA. Inhibition of PKC also completely inhibits PMA mediated caspase-3 activation. On the other hand, caspase 3 inhibition, lessens the PMA induced-effects on the appearance of RBC morphology alterations, although it enhances PMA-mediated effects on nitric oxide (NO) derived metabolites levels. These data demonstrate that PKC-mediated activation of caspase 3 is an integral and essential part of signaling pathway in RBCs, that may be a regulatory factor of RBC mechanical properties, through regulation of NO metabolism.
我们之前已经表明，红细胞（RBCs）的形态改变与内皮型一氧化氮合酶（eNOS）酶活性受损以及随之而来的一氧化氮（NO）衍生代谢产物生成减少相关。在此，我们扩展了之前的观察结果，报告红细胞形态受一系列与蛋白激酶C（PKC）介导的半胱天冬酶3激活相关的特定细胞信号事件调控。在用PKC激活剂佛波醇-12-肉豆蔻酸酯-13-乙酸酯（PMA）处理之前，先用PKC抑制剂白屈菜红碱预处理红细胞，可阻断形态改变的出现以及PMA诱导的硝酸盐和亚硝酸盐水平的持续降低。抑制PKC也完全抑制PMA介导的半胱天冬酶-3激活。另一方面，抑制半胱天冬酶3可减轻PMA对红细胞形态改变出现的诱导作用，尽管它增强了PMA对一氧化氮（NO）衍生代谢产物水平的影响。这些数据表明，PKC介导的半胱天冬酶3激活是红细胞信号通路中不可或缺的重要部分，通过调节NO代谢，它可能是红细胞机械特性的调节因子。
