Mitochondrial oxidative stress induces leaky ryanodine receptor during mechanical ventilation
Mitochondrial oxidative stress induces leaky ryanodine receptor during mechanical ventilation
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DOI：
10.1016/j.freeradbiomed.2019.11.019
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发表时间：
2020-01-01
影响因子：
7.4
通讯作者：
Matecki, Stephan
中科院分区：
文献类型：
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作者：
Dridi, Haikel;Yehya, Mohamad;Matecki, Stephan
理论基础：呼吸机诱发的隔膜功能障碍(VIDD)会增加重症监护患者的发病率和死亡率。虽然VIDD与线粒体氧化应激和钙稳态受损有关，但其潜在的机制仍不清楚。我们假设隔膜线粒体氧化应激引起兰尼定受体(RyR1)/钙释放通道的重塑，导致肌浆网(SR)钙泄漏、蛋白分解和VIDD。方法：在机械通气短时间(6h)和长时间(12h)的小鼠横隔膜中，我们评估了线粒体ROS的产生、线粒体乌头酸酶活性作为线粒体氧化应激的标志、RyR1的重塑和功能、钙依赖的蛋白分解、转化生长因子β1和STAT3途径、肌肉纤维横截面积和横隔膜比力的产生。6‘-二甲基酪氨酸-赖氨酸-苯丙氨酸-氨基转移酶(6’-二甲基酪氨酸-赖氨酸-苯丙氨酸-氨基)2，6‘-二甲基酪氨酸-赖氨酸-苯丙氨酸-氨基转移酶(6’-二甲基酪氨酸-赖氨酸-苯丙氨酸-NH_2)。测定和主要结果：机械通气(MV)6h导致线粒体ROS生成增加，线粒体乌头酸酶活性降低，氧化作用增强，S亚硝化、S-谷胱甘肽和Ser-2844磷酸化，RyR1稳定亚基calstein1耗尽，肌浆网钙外泄增加。通过SS31处理阻止MROS的产生并不影响转化生长因子β1和信号转导通路3的激活，这表明线粒体氧化应激是转化生长因子β1和信号转导通路3的下游通路，而转化生长因子β1和信号转导通路3早期参与了VIDD。SS-31挽救了MV引起的所有其他细胞事件和膈肌收缩功能障碍，而SS20(一种缺乏抗氧化特性的SS31类似物)未能阻止这些细胞事件和收缩功能障碍。在通风12小时中发现了类似的结果。此外，SS31处理阻止了12小时MV后观察到的钙蛋白1活性和横隔膜萎缩。本研究强调，6小时MV期间线粒体氧化应激通过RyR1重塑和横隔膜无力导致SR钙泄漏，而较长时间的MV(12小时)也与钙依赖的蛋白分解增加和横隔膜萎缩有关。
Rationale: Ventilator-induced diaphragm dysfunction (VIDD) increases morbidity and mortality in critical care patients. Although VIDD has been associated with mitochondrial oxidative stress and calcium homeostasis impairment, the underling mechanisms are still unknown. We hypothesized that diaphragmatic mitochondrial oxidative stress causes remodeling of the ryanodine receptor (RyR1)/calcium release channel, contributing to sarcoplasmic reticulum (SR) Ca2+ leak, proteolysis and VIDD.Method: In mice diaphragms mechanically ventilated for short (6 h) and long (12 h) period, we assessed mitochondrial ROS production, mitochondrial aconitase activity as a marker of mitochondrial oxidative stress, RyR1 remodeling and function, Ca2+ dependent proteolysis, TGF beta 1 and STAT3 pathway, muscle fibers cross-sectional area, and diaphragm specific force production, with or without the mitochondrial targeted anti-oxidant peptide d-Arg-2', 6'-dimethyltyrosine-Lys-Phe-NH2 (SS31).Measurements and main results: 6 h of mechanical ventilation (MV) resulted in increased mitochondrial ROS production, reduction of mitochondrial aconitase activity, increased oxidation, S-nitrosylation, S-glutathionylation and Ser-2844 phosphorylation of RyR1, depletion of stabilizing subunit calstabin1 from RyR1, increased SR Ca2+ leak. Preventing mROS production by SS31 treatment does not affect the TGF beta 1 and STAT3 activation, which suggests that mitochondrial oxidative stress is a downstream pathway to TGF beta 1 and STAT3, early involved in VIDD. This is further supported by the fact that SS-31 rescue all the other described cellular events and diaphragm contractile dysfunction induced by MV, while SS20, an analog of SS31 lacking antioxidant properties, failed to prevent these cellular events and the contractile dysfunction.Similar results were found in ventilated for 12 h. Moreover, SS31 treatment prevented calpain1 activity and diaphragm atrophy observed after 12 h of MV.This study emphasizes that mitochondrial oxidative stress during 6 h-MV contributes to SR Ca2+ leak via RyR1 remodeling, and diaphragm weakness, while longer periods of MV (12 h) were also associated with increased Ca2+-dependent proteolysis and diaphragm atrophy.