Ma Qing, Zhang Zhiquan, Shim Jae-Kwang, Venkatraman Talaignair N, Lascola Christopher D, Quinones Quintin J, Mathew Joseph P, Terrando Niccolò, Podgoreanu Mihai V
Systems Modeling of Perioperative Organ Injury Laboratory, Department of Anesthesiology, Duke University, Durham, NC, United States.
Neuroinflammation and Cognitive Outcomes Laboratory, Department of Anesthesiology, Duke University, Durham, NC, United States.
Front Neurosci. 2019 Jun 14;13:608. doi: 10.3389/fnins.2019.00608. eCollection 2019.
Neuroinflammation initiated by damage-associated molecular patterns, including high mobility group box 1 protein (HMGB1), has been implicated in adverse neurological outcomes following lethal hemorrhagic shock and polytrauma. Emergency preservation and resuscitation (EPR) is a novel method of resuscitation for victims of exsanguinating cardiac arrest, shown in preclinical studies to improve survival with acceptable neurological recovery. Sirtuin 3 (SIRT3), the primary mitochondrial deacetylase, has emerged as a key regulator of metabolic and energy stress response pathways in the brain and a pharmacological target to induce a neuronal pro-survival phenotype. This study aims to examine whether systemic administration of an Annexin-A1 bioactive peptide (ANXA1sp) could resolve neuroinflammation and induce sirtuin-3 regulated cytoprotective pathways in a novel rat model of exsanguinating cardiac arrest and EPR. Adult male rats underwent hemorrhagic shock and ventricular fibrillation, induction of profound hypothermia, followed by resuscitation and rewarming using cardiopulmonary bypass (EPR). Animals randomly received ANXA1sp (3 mg/kg, in divided doses) or vehicle. Neuroinflammation (HMGB1, TNFα, IL-6, and IL-10 levels), cerebral cell death (TUNEL, caspase-3, pro and antiapoptotic protein levels), and neurologic scores were assessed to evaluate the inflammation resolving effects of ANXA1sp following EPR. Furthermore, western blot analysis and immunohistochemistry were used to interrogate the mechanisms involved. Compared to vehicle controls, ANXA1sp effectively reduced expression of cerebral HMGB1, IL-6, and TNFα and increased IL-10 expression, which were associated with improved neurological scores. ANXA1sp reversed EPR-induced increases in expression of proapoptotic protein Bax and reduction in antiapoptotic protein Bcl-2, with a corresponding decrease in cerebral levels of cleaved caspase-3. Furthermore, ANXA1sp induced autophagic flux (increased LC3II and reduced p62 expression) in the brain. Mechanistically, these findings were accompanied by upregulation of the mitochondrial protein deacetylase Sirtuin-3, and its downstream targets FOXO3a and MnSOD in ANXA1sp-treated animals. Our data provide new evidence that engaging pro-resolving pharmacological strategies such as Annexin-A1 biomimetic peptides can effectively attenuate neuroinflammation and enhance the neuroprotective effects of EPR after exsanguinating cardiac arrest.
由损伤相关分子模式引发的神经炎症，包括高迁移率族蛋白B1（HMGB1），与致死性失血性休克和多发伤后的不良神经学后果有关。紧急保存与复苏（EPR）是一种针对失血性心脏骤停受害者的新型复苏方法，临床前研究表明其能提高存活率并使神经功能得到可接受的恢复。沉默调节蛋白3（SIRT3）是主要的线粒体去乙酰化酶，已成为大脑中代谢和能量应激反应途径的关键调节因子以及诱导神经元促生存表型的药理学靶点。本研究旨在探讨在一种新型的失血性心脏骤停和EPR大鼠模型中，全身给予膜联蛋白A1生物活性肽（ANXA1sp）是否能解决神经炎症并诱导沉默调节蛋白3调控的细胞保护途径。成年雄性大鼠经历失血性休克和心室颤动，诱导深度低温，随后使用体外循环进行复苏和复温（EPR）。动物随机接受ANXA1sp（3mg/kg，分剂量给药）或赋形剂。评估神经炎症（HMGB1、TNFα、IL-6和IL-10水平）、脑细胞死亡（TUNEL、半胱天冬酶-3、促凋亡和抗凋亡蛋白水平）以及神经学评分，以评估EPR后ANXA1sp的炎症消退作用。此外，采用蛋白质免疫印迹分析和免疫组织化学来探究其中涉及的机制。与赋形剂对照组相比，ANXA1sp有效降低了大脑中HMGB1、IL-6和TNFα的表达，并增加了IL-10的表达，这与神经学评分的改善相关。ANXA1sp逆转了EPR诱导的促凋亡蛋白Bax表达增加和抗凋亡蛋白Bcl-2减少，同时大脑中裂解的半胱天冬酶-3水平相应降低。此外，ANXA1sp诱导了大脑中的自噬流（增加LC3II并降低p62表达）。从机制上讲，这些发现伴随着线粒体蛋白去乙酰化酶沉默调节蛋白3及其下游靶点FOXO3a和锰超氧化物歧化酶（MnSOD）在接受ANXA1sp治疗的动物中的上调。我们的数据提供了新的证据，即采用促消退药理学策略，如膜联蛋白A1模拟肽，可以有效减轻神经炎症并增强失血性心脏骤停后EPR的神经保护作用。
