Mitochondrial PKC-ε deficiency promotes I/R-mediated myocardial injury via GSK3β-dependent mitochondrial permeability transition pore opening.
Mitochondrial PKC-ε deficiency promotes I/R-mediated myocardial injury via GSK3β-dependent mitochondrial permeability transition pore opening.
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线粒体PKC-ε缺乏通过GSK3β依赖性的线粒体渗透性过渡孔口开放来促进I/R介导的心肌损伤。
DOI：
10.1111/jcmm.13121
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发表时间：
2017-09
影响因子：
5.3
通讯作者：
Zhang YE
中科院分区：
文献类型：
--
作者：
Wang S;Zhang F;Zhao G;Cheng Y;Wu T;Wu B;Zhang YE
线粒体裂变与心肌细胞凋亡密切相关，这被认为是缺血/再灌注（I/R）引起的心肌损伤的主要原因之一。在我们之前的工作中，我们证明乙醛脱氢酶-2 (ALDH2) 缺乏会加剧心肌细胞凋亡和心功能障碍。本研究的目的是阐明 ALDH2 缺乏是否会促进 I/R 应激反应中的线粒体损伤和心肌细胞死亡及其潜在机制。主动脉交叉钳夹 45 分钟诱导 I/R 损伤。然后在 ALDH2 敲除 (ALDH2−/−) 和野生型 (WT) 小鼠中松开 24 小时。然后检查心肌梗塞面积、细胞凋亡和心功能。通过蛋白质印迹法测定蛋白激酶 C (PKC) 亚型表达及其线粒体易位、动力相关蛋白 1 (Drp1)、caspase9 和 caspase3 的活性。还检测了 N-乙酰半胱氨酸 (NAC) 或 PKC-δ shRNA 处理对糖原合成酶激酶-3β (GSK-3β) 活性和线粒体通透性转换孔 (mPTP) 打开的影响。结果显示，ALDH2−/− 小鼠心肌梗塞面积增加，心肌细胞凋亡增加，裂解的 caspase9、caspase3 和磷酸化的 Drp1 水平增加。 ALDH2−/− 小鼠的线粒体 PKC-ε 易位低于 WT 小鼠，而 PKC-δ 则相反。进一步的数据表明，线粒体 PKC 亚型比例受细胞活性氧 (ROS) 水平调节，在 I/R 损伤下，NAC 预处理可以逆转这种情况。此外，PKC-ε 抑制导致 caspase9、caspase3 和 Drp1Ser616 激活以响应 I/R 应激。重要的是，磷酸化 GSK-3β（无活性形式）的表达在 ALDH2−/− 小鼠中低于 WT 小鼠，并且两者均因 NAC 预处理而增加。 PKC-δ shRNA 或 NAC 预处理可抑制 I/R 诱导的 GSK-3β 线粒体易位。此外，ALDH2−/− 小鼠在 I/R 后线粒体膜电位 (ΔΨm) 降低，但 GSK-3β 抑制剂 (SB216763) 或 PKC-δ shRNA 可以部分逆转这种情况。总的来说，我们的数据提供了证据，表明异常的 PKC-ε/PKC-δ 比率促进了 Drp1 信号传导、半胱天冬酶级联和 GSK-3β 依赖性 mPTP 打开的激活，从而导致 I/R 应激后 ALDH2−/− 小鼠线粒体损伤触发的心肌细胞凋亡和心肌功能障碍。
Mitochondrial fission is critically involved in cardiomyocyte apoptosis, which has been considered as one of the leading causes of ischaemia/reperfusion (I/R)‐induced myocardial injury. In our previous works, we demonstrate that aldehyde dehydrogenase‐2 (ALDH2) deficiency aggravates cardiomyocyte apoptosis and cardiac dysfunction. The aim of this study was to elucidate whether ALDH2 deficiency promotes mitochondrial injury and cardiomyocyte death in response to I/R stress and the underlying mechanism. I/R injury was induced by aortic cross‐clamping for 45 min. followed by unclamping for 24 hrs in ALDH2 knockout (ALDH2−/−) and wild‐type (WT) mice. Then myocardial infarct size, cell apoptosis and cardiac function were examined. The protein kinase C (PKC) isoform expressions and their mitochondrial translocation, the activity of dynamin‐related protein 1 (Drp1), caspase9 and caspase3 were determined by Western blot. The effects of N‐acetylcysteine (NAC) or PKC‐δ shRNA treatment on glycogen synthase kinase‐3β (GSK‐3β) activity and mitochondrial permeability transition pore (mPTP) opening were also detected. The results showed that ALDH2−/− mice exhibited increased myocardial infarct size and cardiomyocyte apoptosis, enhanced levels of cleaved caspase9, caspase3 and phosphorylated Drp1. Mitochondrial PKC‐ε translocation was lower in ALDH2−/− mice than in WT mice, and PKC‐δ was the opposite. Further data showed that mitochondrial PKC isoform ratio was regulated by cellular reactive oxygen species (ROS) level, which could be reversed by NAC pre‐treatment under I/R injury. In addition, PKC‐ε inhibition caused activation of caspase9, caspase3 and Drp1Ser616 in response to I/R stress. Importantly, expression of phosphorylated GSK‐3β (inactive form) was lower in ALDH2−/− mice than in WT mice, and both were increased by NAC pre‐treatment. I/R‐induced mitochondrial translocation of GSK‐3β was inhibited by PKC‐δ shRNA or NAC pre‐treatment. In addition, mitochondrial membrane potential (∆Ψm) was reduced in ALDH2−/− mice after I/R, which was partly reversed by the GSK‐3β inhibitor (SB216763) or PKC‐δ shRNA. Collectively, our data provide the evidence that abnormal PKC‐ε/PKC‐δ ratio promotes the activation of Drp1 signalling, caspase cascades and GSK‐3β‐dependent mPTP opening, which results in mitochondrial injury‐triggered cardiomyocyte apoptosis and myocardial dysfuction in ALDH2−/− mice following I/R stress.
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影响因子：
20.1
作者：
Hong Z;Kutty S;Toth PT;Marsboom G;Hammel JM;Chamberlain C;Ryan JJ;Zhang HJ;Sharp WW;Morrow E;Trivedi K;Weir EK;Archer SL
通讯作者：
Archer SL
DOI：
10.1073/pnas.191369098
发表时间：
2001-09-25
影响因子：
11.1
作者：
Chen, L;Hahn, H;Mochly-Rosen, D
通讯作者：
Mochly-Rosen, D
影响因子：
20.1
作者：
Churchill, EN;Murriel, CL;Szweda, LI
通讯作者：
Szweda, LI
影响因子：
37.8
作者：
Inagaki, K;Begley, R;Mochly-Rosen, D
通讯作者：
Mochly-Rosen, D
影响因子：
5
作者：
Churchill, Eric N.;Disatnik, Marie-Helene;Mochly-Rosen, Daria
通讯作者：
Mochly-Rosen, Daria