SH2B1 protects cardiomyocytes from ischemia/reperfusion injury via the activation of the PI3K/AKT pathway
SH2B1 protects cardiomyocytes from ischemia/reperfusion injury via the activation of the PI3K/AKT pathway
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SH2B1 通过激活 PI3K/AKT 通路保护心肌细胞免受缺血/再灌注损伤
DOI：
10.1016/j.intimp.2019.105910
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发表时间：
2020-06-01
影响因子：
5.6
通讯作者：
Chen Man-hua
中科院分区：
文献类型：
--
作者：
Guo Xin;Li Xu-yong;Chen Man-hua
背景资料：细胞凋亡、反应性氧化应激（ROS）和炎症反应是心肌缺血/再灌注（I/R）损伤（MIRI）的重要机制。我们的前期研究和其他研究已经证实src同源2（SH 2）B衔接蛋白1（SH 2 B1）参与了缺血性损伤和心肌肥厚，而SH 2 B1参与MIRI及其机制尚不清楚。方法：本研究采用结扎冠状动脉前降支30 min再灌注24 h的方法建立MIRI动物模型，原代培养的心肌细胞缺氧2 h复氧4 h（H/R）模拟MIRI。在体内和体外MIRI之前，将编码SH 2B 1或GFP的腺病毒预转染到心肌中。系统评价心肌损伤、心功能、细胞凋亡、活性氧及炎症反应。结果：H/R或I/R可显著降低心肌细胞SH 2B 1的表达，并导致细胞存活和功能受损，而腺病毒介导的SH 2B 1上调可逆转上述情况。同时，I/R和H/R升高的炎症、凋亡和ROS也通过SH 2B 1上调而减轻。SH 2B 1对H/R损伤心肌细胞的保护作用机制可能与其激活PI 3 K/AKT通路有关。通过药物抑制剂LY 294002阻断PI 3 K/AKT，SH 2B 1的抗H/R能力受到抑制。结论：SH 2B 1通过PI 3 K/AKT依赖的途径抑制MIRI心肌细胞的炎症、凋亡和ROS的产生。它可能为MIRI的治疗提供新的靶点。
Background: Apoptosis, reactive oxidative stress (ROS) and inflammation act as the pivotal pathogenesis of myocardial ischemia/reperfusion (I/R) injury (MIRI). Our prior study and other investigation have demonstrated the participations of src homology 2 (SH2) B adaptor protein 1 (SH2B1) in ischemic injury and cardiac hypertrophy; whereas, the involvements of SH2B1 in MIRI and underlying mechanisms are completely unknown.Method: In present study, MIRI model in vivo was induced by 30 min of ligation of LAD coronary artery and 24 h of reperfusion, and primary cultured cardiomyocytes were challenged with 2 h of hypoxia followed by 4 h of reoxygenation (H/R) to mimic MIRI in vitro. Adenovirus encoding for SH2B1 or GFP were pre-transfected into myocardium prior to MIRI both in vivo and in vitro. The myocardial damage, cardiac function, apoptosis, ROS and inflammation were evaluated systematically. Immunofluorescence staining and western blotting were alternatively performed to detect protein expression.Results: The results exhibited that H/R or I/R significantly reduced SH2B1 in cardiomyocytes, followed by impaired cell survival and function, which were strongly reversed after the adenovirus-mediated SH2B1 upregulation. Meanwhile, I/R- and H/R-elevated inflammation, apoptosis and ROS were also alleviated by SH2B1 up-regulation. A mechanistic study suggested that the protective contributions of SH2B1 on H/R-suffered cardiomyocytes were based on the activation of the PI3K/AKT pathway. The abolishment of the PI3K/AKT via a pharmacological inhibitor (LY294002) repressed anti-H/R capabilities of SH2B1.Conclusion: Therefore, SH2B1 prevents cardiomyocytes from inflammation, apoptosis and ROS in MIRI partially through the PI3K/AKT-dependent avenues. It may provide a novel therapeutic target for the treatment of MIRI.