Novel Mitochondrial Mechanisms of Ethanol-Induced Cardiac Injury in Aged Females

老年女性乙醇诱发心脏损伤的线粒体新机制

• 批准号: 7944093
• 负责人: DONNA HOPE KORZICK
• 金额: $ 50.51万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7944093
• 关键词: Address; Adult; Age; Aging; Alcohol consumption; Alcohol dependence; Alcoholism; Alcohols; Amino Acid Sequence; Animal Model; Apoptotic; Arts; Biochemical; Biological Assay; Cardiac; Cardiovascular system; Cause of Death; Cell Death; Cell Survival; Centers for Disease Control and Prevention (U.S.); Centrifugation; Cessation of life; Chronic; Clinical Research; Clinical Trials; Complex; Computer software; Development; Epidemiology; Estrogen Receptors; Estrogens; Ethanol; Exhibits; Female; Functional disorder; Future; Gel; Gender; Glean; Heart; Heart Diseases; Heart failure; Heavy Drinking; Hypertension; In Vitro; Individual; Infarction; Injury; Ischemia; Ischemic Preconditioning; Label; Maintenance; Metabolism; Methodology; Mitochondria; Mitochondrial Proteins; Morbidity - disease rate; Myocardial Infarction; Myocardium; Necrosis; Ontology; Operative Surgical Procedures; Ovary; Oxidative Stress; Pathway interactions; Peptide Sequence Determination; Phenotype; Physiological; Play; Population; Post-Translational Protein Processing; Postmenopause; Predisposition; Production; Proteins; Proteomics; Public Health; Rattus; Reactive Oxygen Species; Recovery; Relative (related person); Reperfusion Injury; Reperfusion Therapy; Respiration; Role; Sampling; Sex Characteristics; Shapes; Signal Pathway; Signal Transduction; Technology; Testing; Therapeutic Intervention; Time; Toxic effect; Western Blotting; Woman; Women' s Role; abstracting; age effect; age related; aged; alcohol effect; cell growth regulation; chronic alcohol ingestion; design; drinking; in vivo; insight; interest; juvenile animal; mature animal; men; middle age; mortality; multiple reaction monitoring; new therapeutic target; novel; novel strategies; pressure; problem drinker; protein expression; public health relevance; research study; response; senescence; sex

DESCRIPTION (provided by applicant): ABSTRACT Sex differences in the cardiovascular response to alcoholism exist such that women are apparently more sensitive to the toxic effects of alcohol vs men, although the mechanisms which underlie this phenomenon are poorly understood and have not been systematically investigated. Greater vulnerability to alcoholic heart disease in women is exacerbated by evidence indicating that post-menopausal women have higher mortality rates after myocardial infarction vs aged men, and it is unclear whether the effects of aging and estrogen (E2) deficiency are additive with regard to ethanol-induced cardiac ischemia/reperfusion (I/R) injury. Because of the pivotal role played by the mitochondria in the maintenance of cell survival and cardioprotection, it is logical that ethanol and age-associated increases in I/R injury might arise from altered responses in the mitochondrial subproteome. To address this problem, we will employ in vitro isolated heart and in vivo mitochondrial respiration studies in conjunction with the newly emerging iTRAQ 8plex proteomics approach, LC MS/MS and PANTHER ontological analysis to characterize alterations in mitochondrial signaling occurring with aging and E2 deficiency in the female rat myocardium following chronic ethanol ingestion. Of particular interest are adaptive changes in proteins associated with metabolism, oxidative stress, and cell death. Specific aims are as follows: 1) chronic ethanol ingestion will exacerbate age-related increases in I/R injury in association with reduced mitochondrial targeting of known cardioprotective signals and estrogen receptor (ER) subtypes, and 2) chronic ethanol ingestion will exacerbate the effects of E2 deficiency on age dependent disruptions in protective mitochondrial signaling. Infarct size and apoptotic cell death will be assessed in Langendorff-perfused hearts following global I/R in adult and aged ovary-intact and gonadectomized female rats; respiration studies will be performed in isolated mitochondria. Western blotting and qRT- PCR will be employed for mitochondrial ER subtype targeting and downstream signals; iTRAQ and multiple reaction monitoring (MRM) for protein changes and post-translational modifications of ERs. The high throughput, targeted iTRAQ proteomics approach we describe here will allow us to provide critical and novel mechanistic insight, in a relatively short time, on the role of female sex in the development of alcohol related cardiac dysfunction in aged and adult rats. Moreover, information gleaned from the proposed studies should provide strong, direct support for new therapeutic targets for treating alcoholic heart disease in postmenopausal women, as well as provide impetus and rationale for the design of future experimental studies. PUBLIC HEALTH RELEVANCE: Current experimental evidence supports a J- or U-shaped association between alcohol consumption and heart attack and/or high blood pressure. As such, while light-to-moderate drinking has been associated with protection from heart disease, heavy drinking is associated with the development of heart failure, which remains a major public health crisis and a leading cause of morbidity and mortality in aged men and women. The CDC has most recently estimated that ~75,000 deaths per year can be attributed to the harmful effects of alcohol. Moreover, nearly 33% of chronic alcohol-dependent individuals exhibit cardiac dysfunction, which often becomes manifest as an alcohol-specific heart failure. Interestingly, women appear to more sensitive to the toxic effects of alcohol on the heart when compared to men, even though total alcohol consumption appears to be less in alcoholic women vs men. It is also important to note that heart disease remains the leading cause of death in post-menopausal women, with mortality rates surpassing those of age-matched men. The specific mechanisms which produce this gender-specific cardiac dysfunction and enhanced susceptibility to ischemic injury in the aged female heart following chronic ethanol ingestion have not been directly examined, and the studies proposed herein will fill this information gap. We propose a comprehensive and novel approach that will identify proteins that comprise the functional sequelae of complex cardiac phenotypes of chronic ethanol ingestion, and will further characterize possible targets for therapeutic intervention in the aged female population.

描述（由申请人提供）：摘要对酗酒的心血管反应存在性别差异，因此女性显然比男性对酒精的毒性作用更敏感，尽管人们对这种现象背后的机制知之甚少，也没有进行系统研究。有证据表明，与老年男性相比，绝经后女性心肌梗死后的死亡率更高，这加剧了女性更容易患酒精性心脏病，而且目前尚不清楚衰老和雌激素 (E2) 缺乏的影响是否与乙醇相加-诱发的心脏缺血/再灌注（I/R）损伤。由于线粒体在维持细胞存活和心脏保护方面发挥着关键作用，因此乙醇和年龄相关的 I/R 损伤增加可能是由线粒体亚蛋白质组反应的改变引起的，这是合乎逻辑的。为了解决这个问题，我们将采用体外离体心脏和体内线粒体呼吸研究，结合新兴的 iTRAQ 8plex 蛋白质组学方法、LC MS/MS 和 PANTHER 本体分析来表征衰老和 E2 缺乏引起的线粒体信号变化。慢性摄入乙醇后雌性大鼠的心肌。特别令人感兴趣的是与新陈代谢、氧化应激和细胞死亡相关的蛋白质的适应性变化。具体目标如下：1) 长期摄入乙醇将加剧与年龄相关的 I/R 损伤增加，这与已知心脏保护信号和雌激素受体 (ER) 亚型的线粒体靶向减少有关，2) 长期摄入乙醇将加剧这种影响E2 缺乏对年龄依赖性线粒体保护信号破坏的影响。在成年和老年卵巢完整和性腺切除的雌性大鼠的整体 I/R 后，将在 Langendorff 灌注的心脏中评估梗死面积和凋亡细胞死亡；呼吸研究将在分离的线粒体中进行。 Western blotting 和 qRT-PCR 将用于线粒体 ER 亚型靶向和下游信号； iTRAQ 和多反应监测 (MRM)，用于检测蛋白质变化和 ER 翻译后修饰。我们在此描述的高通量、靶向 iTRAQ 蛋白质组学方法将使我们能够在相对较短的时间内提供关于雌性在老年和成年大鼠酒精相关心脏功能障碍发展中的作用的关键和新颖的机制见解。此外，从拟议研究中收集的信息应该为治疗绝经后妇女酒精性心脏病的新治疗目标提供强有力的直接支持，并为未来实验研究的设计提供动力和理由。 公共卫生相关性：当前的实验证据支持饮酒与心脏病和/或高血压之间存在 J 形或 U 形关联。因此，虽然轻度至中度饮酒与预防心脏病有关，但大量饮酒与心力衰竭的发生有关，心力衰竭仍然是一个重大的公共卫生危机，也是老年男性和女性发病和死亡的主要原因。疾病预防控制中心 (CDC) 最近估计，每年约有 75,000 人死亡可归因于酒精的有害影响。此外，近 33% 的慢性酒精依赖者表现出心脏功能障碍，这通常表现为酒精特异性心力衰竭。有趣的是，与男性相比，女性似乎对酒精对心脏的毒性作用更敏感，尽管酗酒女性的饮酒总量似乎少于男性。还需要注意的是，心脏病仍然是绝经后女性死亡的主要原因，其死亡率超过了同龄男性。长期摄入乙醇后老年女性心脏产生这种性别特异性心脏功能障碍和对缺血性损伤的易感性增强的具体机制尚未得到直接研究，本文提出的研究将填补这一信息空白。我们提出了一种全面而新颖的方法，该方法将鉴定构成慢性乙醇摄入的复杂心脏表型的功能性后遗症的蛋白质，并将进一步确定老年女性群体治疗干预的可能目标。