Mitochondrial Ca-sensitive K channel-induced superoxide and cardiac protection

线粒体钙敏感钾通道诱导的超氧化物和心脏保护

• 批准号: 8208049
• 负责人: DAVID F STOWE
• 金额: $ 33.75万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8208049
• 关键词: Agonist; Anions; Apoptosis; Bioenergetics; Cardiac; Cardiac Myocytes; Cavia; Cells; Characteristics; Charge; Complement; Complex; Consumption; Coronary Arteriosclerosis; Coupled; Cytoprotection; Data; Drug Delivery Systems; Electron Transport; Electrons; Event; Exposure to; Feedback; Generations; Glutathione; Health; Heart; Heart Atrium; Homeostasis; Hydrogen Peroxide; In Situ; Infarction; Inner mitochondrial membrane; Ischemia; Lead; Link; Lipid Bilayers; Mass Fragmentography; Mass Spectrum Analysis; Measures; Mechanics; Mediating; Membrane; Membrane Potentials; Memory; Metabolic; Mitochondria; Mitochondrial Matrix; Modeling; Molecular; Muscle Cells; NADH; Necrosis; Oxidation-Reduction; Pathway interactions; Patients; Peptides; Perfusion; Pharmaceutical Preparations; Phosphotransferases; Physiologic pulse; Play; Potassium Channel; Proteins; Proton Pump; Public Health; Reaction; Reactive Oxygen Species; Regulation; Reperfusion Injury; Reperfusion Therapy; Research; Respiration; Respiratory Chain; Role; Scheme; Source; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Stimulus; Superoxides; Swelling; Techniques; Testing; Tissues; Two-Dimensional Gel Electrophoresis; Vascular Smooth Muscle; Western Blotting; catalase; cell injury; cell type; conditioning; improved; improved functioning; indexing; inhibitor/antagonist; mimetics; novel strategies; practical application; preconditioning; protective effect; respiratory; skills; tool; voltage

DESCRIPTION (provided by applicant): Pharmacologic preconditioning (PPC) is an approach to limit ischemia reperfusion (IR) injury that does not require prior brief ischemia or the presence of the drug once ischemia occurs. KATP channel agonists, per se, induce PPC via memory pathways that lead to cardioprotection assessed by better tissue perfusion, improved metabolic and mechanical function, fewer dysrhythmias and reduced infarct size. Limits to our understanding of features and mechanisms of PPC have impeded optimal utilization of this potent phenomenon to protect against cardiac IR injury. Particularly lacking is an understanding of the role of the mitochondrion in PPC. Big (B) conductance Ca2+ -sensitive K+ channels (BKCa) are also present in cardiac cell inner mitochondrial membrane (IMM) and appear to mediate cardioprotection. We have evidence of protection also by small conductance (S) KCa channels in the IMM. We propose that drug -induced K+ entry into the mitochondrial (m) matrix alters bioenergetics in a way that increases electron leak to induce an increase in reactive oxygen species (ROS) required to trigger downstream protective effects. Preliminary results indicate that protective effects of mKCa channel opening are blocked by dismutation of the superoxide radical. A unifying hypothesis for the initiating mechanism of PPC may be a matrix K+ influx -induced ROS generation. We will a) examine in guinea pig isolated cardiac mitochondria the bioenergetic mechanisms initiated by matrix K+ influx that lead to ROS generation and b) determine the specific ROS responsible for triggering PPC in guinea pig isolated hearts. In addition we will c) identify these channels by Western blots, 2D gel electrophoresis and MALDI- TOF and LIT SNCE mass spectrometry and characterize these channels in artificial lipid bilayers. We will use the best techniques, measures (mitochondrial respiration, cytosolic and mCa2+, NADH, mFAD, mpH, IMM potential, and several ROS) and drugs available to search for the factors, the sequence of events, and the specific ROS that initiate PPC. PUBLIC HEALTH RELEVANCE: These studies will result in a better understanding of the regulation of mitochondrial bioenergetic function by Ca2+, K+, and ROS, and selection of the mitochondrion as a target for pharmacologic manipulation. This research should lead to the practical application of mitochondrial-targeted drugs to prophylactically treat patients with coronary artery disease using novel approaches.

描述（由申请人提供）：药物预处理（PPC）是一种限制缺血再灌注（IR）损伤的方法，不需要事先简短缺血或一旦发生缺血就存在药物。 KATP通道激动剂本身，通过记忆途径诱导PPC，导致通过更好的组织灌注，改善的代谢和机械功能评估心脏保护，减少障碍和梗塞大小。我们对PPC特征和机制的理解的限制阻碍了这种有效现象的最佳利用，以防止心脏IR损伤。特别缺乏的是了解线粒体在PPC中的作用。大（B）电导Ca2+敏感的K+通道（BKCA）也存在于心脏细胞内线粒体膜（IMM）中，并且似乎介导心脏保护。我们也有少量电导率在IMM中提供保护的证据。我们提出，药物诱导的K+进入线粒体（M）基质以增加电子泄漏的方式改变了生物能，以诱导触发下游保护作用所需的活性氧（ROS）增加。初步结果表明，MKCA通道开口的保护作用被超氧化物自由基的拆除所阻断。 PPC启动机制的统一假设可能是基质K+涌入引起的ROS产生。我们将a）在豚鼠分离的心脏线粒体中检查由基质K+涌入引发的生物能机制，导致ROS产生和b）确定负责在豚鼠分离的心脏中触发PPC的特定ROS。此外，我们将c）通过蛋白质印迹，2D凝胶电泳和Maldifof和LIT SNCE质谱识别这些通道，并在人工脂质双层中表征这些通道。我们将使用最佳技术，措施（线粒体呼吸，胞质和MCA2+，NADH，MFAD，MPH，MPH，IMM潜力以及几种ROS）以及可用于寻找因素，事件的顺序以及启动PPC的特定ROS的药物。公共卫生相关性：这些研究将更好地理解Ca2+，K+和ROS对线粒体生物能功能的调节，并选择线粒体作为药理操作的目标。这项研究应导致用新方法使用线粒体靶向药物在预防性治疗冠状动脉疾病的患者中实际应用。

项目成果

Protection against cardiac injury by small Ca(2+)-sensitive K(+) channels identified in guinea pig cardiac inner mitochondrial membrane.

• DOI: 10.1016/j.bbamem.2012.08.031
• 发表时间: 2013-02
• 期刊: BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
• 影响因子: 3.4
• 作者: Stowe, David F.;Gadicherla, Ashish K.;Zhou, Yifan;Aldakkak, Mohammed;Cheng, Qunli;Kwok, Wai-Meng;Jiang, Ming Tao;Heisner, James S.;Yang, MeiYing;Camara, Amadou K. S.
• 通讯作者: Camara, Amadou K. S.

Reduced mitochondrial Ca2+ loading and improved functional recovery after ischemia-reperfusion injury in old vs. young guinea pig hearts.

与年轻豚鼠心脏相比，减少了线粒体 Ca2 负荷并改善了缺血再灌注损伤后的功能恢复。

• DOI: 10.1152/ajpheart.00533.2011
• 发表时间: 2012
• 期刊: American journal of physiology. Heart and circulatory physiology
• 作者: Rhodes,SamhitaS;Camara,AmadouKS;Heisner,JamesS;Riess,MatthiasL;Aldakkak,Mohammed;Stowe,DavidF
• 通讯作者: Stowe,DavidF

Reversible blockade of complex I or inhibition of PKCβ reduces activation and mitochondria translocation of p66Shc to preserve cardiac function after ischemia.

• DOI: 10.1371/journal.pone.0113534
• 发表时间: 2014
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Yang M;Stowe DF;Udoh KB;Heisner JS;Camara AK
• 通讯作者: Camara AK

Damage to mitochondrial complex I during cardiac ischemia reperfusion injury is reduced indirectly by anti-anginal drug ranolazine.

• DOI: 10.1016/j.bbabio.2011.11.021
• 发表时间: 2012-03
• 期刊: BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS
• 影响因子: 4.3
• 作者: Gadicherla, Ashish K.;Stowe, David F.;Antholine, William E.;Yang, Meiying;Camara, Amadou K. S.
• 通讯作者: Camara, Amadou K. S.

Extra-matrix Mg2+ limits Ca2+ uptake and modulates Ca2+ uptake-independent respiration and redox state in cardiac isolated mitochondria.

• DOI: 10.1007/s10863-013-9500-5
• 发表时间: 2013-06
• 期刊: JOURNAL OF BIOENERGETICS AND BIOMEMBRANES
• 影响因子: 3
• 作者: Boelens, Age D.;Pradhan, Ranjan K.;Blomeyer, Christoph A.;Camara, Amadou K. S.;Dash, Ranjan K.;Stowe, David F.
• 通讯作者: Stowe, David F.