Reversible blockade of complex I or inhibition of PKCβ reduces activation and mitochondria translocation of p66Shc to preserve cardiac function after ischemia.
Reversible blockade of complex I or inhibition of PKCβ reduces activation and mitochondria translocation of p66Shc to preserve cardiac function after ischemia.
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DOI：
10.1371/journal.pone.0113534
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Camara AK
中科院分区：
文献类型：
--
作者：
Yang M;Stowe DF;Udoh KB;Heisner JS;Camara AK
过量的线粒体活性氧（mROS）在心肌缺血再灌注（IR）损伤中起重要作用。P66 Shc是ShcA衔接蛋白家族的剪接变体，通过氧化还原的细胞色素c产生H2 O2来增强mROS的产生。消融p66 Shc可保护IR损伤，但尚不清楚p66 Shc在心肌缺血和/或再灌注期间是否以及何时被激活，以及减弱复合物I电子传递或失活PKCβ是否改变IR期间p66 Shc的激活与心脏保护相关。离体豚鼠心脏灌流，并在有或没有异戊巴比妥（复合物I阻断剂）或hispidin（PKCβ抑制剂）的情况下进行缺血和再灌注时间的增加。测定p66 Shc在丝氨酸36位的磷酸化以及线粒体和细胞质中p66 Shc的水平。在线监测缺血前、缺血中、缺血后心功能指标和氧化还原状态。再灌注120分钟后，在一些心脏中评估血管大小。在缺血20和30 min后再灌注期间，p66 Shc磷酸化及其向线粒体的移位增加，但仅在缺血期间或在5或10 min缺血20 min再灌注期间不增加。相应地，在这些缺血和再灌注期间，胞质p66 Shc水平下降。异戊巴比妥或hispidin减少缺血30 min和再灌注20 min诱导的p66 Shc磷酸化及其线粒体易位。异戊巴比妥或hispidin降低p66 Shc的磷酸化导致更好的功能恢复和更少的再灌注期间的梗死。我们的研究结果表明，IR激活p66 Shc，可逆阻断复合物I的电子传递，或抑制PKCβ的激活，减少p66 Shc的激活和易位，减少IR损伤。这些观察结果支持一种新的潜在的治疗干预心脏IR损伤。
Excess mitochondrial reactive oxygen species (mROS) play a vital role in cardiac ischemia reperfusion (IR) injury. P66Shc, a splice variant of the ShcA adaptor protein family, enhances mROS production by oxidizing reduced cytochrome c to yield H2O2. Ablation of p66Shc protects against IR injury, but it is unknown if and when p66Shc is activated during cardiac ischemia and/or reperfusion and if attenuating complex I electron transfer or deactivating PKCβ alters p66Shc activation during IR is associated with cardioprotection. Isolated guinea pig hearts were perfused and subjected to increasing periods of ischemia and reperfusion with or without amobarbital, a complex I blocker, or hispidin, a PKCβ inhibitor. Phosphorylation of p66Shc at serine 36 and levels of p66Shc in mitochondria and cytosol were measured. Cardiac functional variables and redox states were monitored online before, during and after ischemia. Infarct size was assessed in some hearts after 120 min reperfusion. Phosphorylation of p66Shc and its translocation into mitochondria increased during reperfusion after 20 and 30 min ischemia, but not during ischemia only, or during 5 or 10 min ischemia followed by 20 min reperfusion. Correspondingly, cytosolic p66Shc levels decreased during these ischemia and reperfusion periods. Amobarbital or hispidin reduced phosphorylation of p66Shc and its mitochondrial translocation induced by 30 min ischemia and 20 min reperfusion. Decreased phosphorylation of p66Shc by amobarbital or hispidin led to better functional recovery and less infarction during reperfusion. Our results show that IR activates p66Shc and that reversible blockade of electron transfer from complex I, or inhibition of PKCβ activation, decreases p66Shc activation and translocation and reduces IR damage. These observations support a novel potential therapeutic intervention against cardiac IR injury.
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影响因子：
64.5
作者：
Giorgio, M;Migliaccio, E;Pelicci, PG
通讯作者：
Pelicci, PG
影响因子：
5.5
作者：
Camara, Amadou K. S.;Aldakkak, Mohammed;Stowe, David F.
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DOI：
10.1152/ajpheart.00711.2002
发表时间：
2003-02-01
影响因子：
4.8
作者：
Kevin, LG;Camara, AKS;Stowe, DF
通讯作者：
Stowe, DF
影响因子：
5.5
作者：
Aldakkak, Mohammed;Stowe, David F.;Camara, Amadou K. S.
通讯作者：
Camara, Amadou K. S.
影响因子：
4.8
作者：
Chen, Q;Vazquez, EJ;Lesnefsky, EJ
通讯作者：
Lesnefsky, EJ