Departamento de Biomedicina Cardiovascular, Instituto Nacional de Cardiología Ignacio Chávez, Juan Badiano No. 1, Colonia Sección XVI, Mexico City 14080, Mexico.
Área de Medicina Experimental y Traslacional, Departamento de Ciencias de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Mexico City 14080, Mexico.
Cells. 2023 Feb 2;12(3):492. doi: 10.3390/cells12030492.
Caveolae-associated signaling toward mitochondria contributes to the cardioprotective mechanisms against ischemia-reperfusion (I/R) injury induced by ischemic postconditioning. In this work, we evaluated the role that the actin-cytoskeleton network exerts on caveolae-mitochondria communication during postconditioning. Isolated rat hearts subjected to I/R and to postconditioning were treated with latrunculin A, a cytoskeleton disruptor. Cardiac function was compared between these hearts and those exposed only to I/R and to the cardioprotective maneuver. Caveolae and mitochondria structures were determined by electron microscopy and maintenance of the actin-cytoskeleton was evaluated by phalloidin staining. Caveolin-3 and other putative caveolae-conforming proteins were detected by immunoblot analysis. Co-expression of caveolin-3 and actin was evaluated both in lipid raft fractions and in heart tissue from the different groups. Mitochondrial function was assessed by respirometry and correlated with cholesterol levels. Treatment with latrunculin A abolishes the cardioprotective postconditioning effect, inducing morphological and structural changes in cardiac tissue, reducing F-actin staining and diminishing caveolae formation. Latrunculin A administration to post-conditioned hearts decreases the interaction between caveolae-forming proteins, the co-localization of caveolin with actin and inhibits oxygen consumption rates in both subsarcolemmal and interfibrillar mitochondria. We conclude that actin-cytoskeleton drives caveolae signaling to mitochondria during postconditioning, supporting their functional integrity and contributing to cardiac adaption against reperfusion injury.
陷窝小体相关信号向线粒体的传递有助于缺血后处理减轻缺血再灌注(I/R)损伤的心脏保护机制。在这项工作中,我们评估了肌动蛋白细胞骨架网络在缺血后处理期间陷窝小体-线粒体通讯中的作用。缺血再灌注和后处理的分离大鼠心脏用细胞骨架破坏剂 latrunculin A 处理。比较这些心脏与仅暴露于 I/R 和心脏保护操作的心脏之间的心脏功能。通过电子显微镜确定陷窝小体和线粒体的结构,并通过鬼笔环肽染色评估肌动蛋白细胞骨架的维持情况。通过免疫印迹分析检测窖蛋白-3 和其他假定的陷窝小体形成蛋白。在不同组的心外膜脂质筏和组织中评估窖蛋白-3 和肌动蛋白的共表达。通过呼吸测定法评估线粒体功能,并与胆固醇水平相关联。用 latrunculin A 处理会消除缺血后处理的心脏保护作用,导致心脏组织的形态和结构发生变化,减少 F-肌动蛋白染色并减少陷窝小体的形成。给予 latrunculin A 后处理心脏会减少陷窝小体形成蛋白之间的相互作用、窖蛋白与肌动蛋白的共定位,并抑制亚肌膜和肌间线粒体的耗氧量。我们得出结论,肌动蛋白细胞骨架在缺血后处理期间驱动陷窝小体向线粒体传递信号,支持它们的功能完整性,并有助于心脏适应再灌注损伤。