CREG protects from myocardial ischemia/reperfusion injury by regulating myocardial autophagy and apoptosis
CREG protects from myocardial ischemia/reperfusion injury by regulating myocardial autophagy and apoptosis
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CREG通过调节心肌自噬和凋亡保护心肌缺血/再灌注损伤
DOI：
10.1016/j.bbadis.2016.11.015
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发表时间：
2017-08-01
影响因子：
6.2
通讯作者：
Han, Yaling
中科院分区：
文献类型：
--
作者：
Song, Haixu;Yan, Chenghui;Han, Yaling
目的：人E1 A刺激基因的细胞阻遏物（CREG）是一种分泌型糖蛋白，可调节组织和细胞的稳态，并具有抗心脏纤维化的作用，这表明CREG对心肌细胞慢性损伤具有潜在的保护作用。然而，CREG在心肌组织急性损伤中的作用尚不清楚，本研究旨在探讨CREG在心肌缺血/再灌注（MI/R）损伤中的作用，并阐明其作用机制。野生型Creg（Creg（+）/（+）），杂合Creg（Creg（+）/（-））小鼠和输注重组03 mg/kg.d CREG蛋白预处理的小鼠（reCreg（+）/（+））组行左冠状动脉缺血30 min再灌注24 h。采用氯化三苯四氮唑（TTC）-伊文思蓝（Evan 'sBlue-Triphenyl-TetrazoliumChloride，TTC）比色法和超声心动图法观察CREG对MI/R小鼠心肌缺血再灌注损伤的影响。通过体外培养心肌细胞进一步研究其作用机制。我们的研究结果表明，小鼠心肌梗死/再灌注后，小鼠心脏CREG蛋白的水平显着下降。与Creg（+）/（+）小鼠相比，Creg（+）/（-）小鼠再灌注后2 h梗死面积更大，Ml/R损伤后28天心功能更差。而reCreg（+）/（+）小鼠即使在MI/R损伤后仍能维持高水平的CREG，并显著缩小梗死范围和改善心功能。在Creg（+）/（-）小鼠中，心肌自噬功能障碍的特征在于LOA和p62的积聚，而凋亡细胞数量增加通过切割的caspase-3印迹和TUNEL染色检测。相反，在reCreg（+）/（+）小鼠中检测到细胞凋亡减少和自噬激活。此外，氯喹，一种自噬黑剂，被用来证明重组CREG保护心肌细胞对凋亡介导的激活自噬在体内和体外。结论：CREG通过激活溶酶体自噬，对心肌缺血再灌注损伤诱导的心肌细胞凋亡具有保护作用。(C)© 2016 Elsevier B. V.版权所有。
Aims: Human cellular repressor of E1A-stimulated genes (CREG) is a secreted glycoprotein that regulates tissue and cell homeostasis and has been shown to antagonize heart fibrosis, which indicates a potential protective effect of CREG against cardiomyocyte chronic damage. However, little is known about the role of CREG in myocardial tissue acute injury, in this study, we aimed to investigate the role of CREG in myocardial ischemia/reperfusion (MI/R) injury and clarify the mechanism of action.Methods and results: Wild-type Creg (Creg (+)/(+)), heterozygous Creg (Creg(+)/(-)) mice and mice pretreated with infusion of recombinant 03 mg/kg.d CREG protein (reCreg(+)/(+)) were subjected to 30 min of left ascending coronary ischemia and 24 h of reperfusion. Evan's Blue-triphenyl- tetrazolium chloride (TTC) solution and echocardiography analysis were used to evaluate the effects of CREG on MI/R mice. The underlying mechanisms were further determined by cultured myocardial cells in vitro. Our findings revealed that the level of CREG protein in mouse hearts was significantly decreased after mice were subjected to MI/R. Moreover, Creg(+)/(-) mice had larger infarction size 2 h after reperfusion and worse cardiac function 28 days after Ml/R injury compared to that in Creg(+)/(+) mice. However, reCreg(+)/(+) mice could maintain CREG at a high level even after MI/R injury, and mitigated infarction size and improved cardiac function significantly. In Creg(+)/(-) mice, myocardial autophagy was dysfunctional characterized by accumulation of LOA and p62, while apoptotic cell number increase was detected by cleaved caspase-3 blotting and TUNEL staining. Conversely, decreased apoptosis and activated autophagy were detected in reCreg(+)/(+) mice. Furthermore, chloroquine, a kind of autophagy blacker, was used to demonstrate recombinant CREG protected cardiomyocytes against apoptosis mediated by activating autophagy both in vivo and in vitro. Finally, we found CREG was involved into lysosomal protein transfer and improve cellular autophagy.Conclusion: CREG protects heart against MI/R injury -induced cardiomyocytes apoptosis by activating lysosomal autophagy.This article is part of a Special Issue entitled: Genetic and epigenetic control of heart failure - edited by Jun Ren and Megan Yingmei Zhang. (C) 2016 Elsevier B.V. All rights reserved.