Ion Gradients And Metabolic Energy In Animal Tissue

动物组织中的离子梯度和代谢能

• 批准号: 7317738
• 负责人: richard l veech
• 金额: --
• 依托单位: NATIONAL INSTITUTE ON ALCOHOL ABUSE AND ALCOHOLISM
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7317738
• 关键词: Ion; Gradients; Metabolic; Energy; Animal

Summary: The energy of the gradients of the nine major inorganic ions in working perfused heart are in near equilibrium with each other, the electrical potential between extra- and intracellular phase and the delta G of ATP hydrolysis [Masuda T et al, J Biol Chem 1990;265:20321-34]. The metabolism of ethanol increases the resting electrical potential of hepatocytes from -28 to -40 mV [Veech RL et al, Alch Clin Exp Res 1994;18:1040-56]. Previously, we showed that merely changing the substrate available altered the delta G of ATP hydrolysis in heart and improved the efficiency of hydraulic work of heart by 28% [Kashiwaya Y et al, Am J Cardiol 1997;80:50A-64A]. Since injuries of any sort induce a stereotypic change in cellular ionic distributions wherein the cell gains Na+, loses K+ and swells, these stereotypic changes of injury can possibly be reversed by simple changes in the compositions of fluids administered to victims of injury or burns. We went on to show that the gradients of all nine common inorganic ions between intra and extracellular phases of a cell form a Donnan near equilibrium system relating the electrical potential between the phases to the energy of ATP hydrolysis [Veech RL et al, IUBMB Life 2002;54:241-52]. As a result of these studies and our earlier studies criticizing the composition of existing parenteral fluids [Veech RL, Am J Clin Nutr 1986;44:519-51] a panel convened by the Academy of Medicine, recommended that investigation of the feasibility of making new resuscitation fluids be initiated [Fluid Resuscitation, Natl Acad Press, Wash DC, 1999]. We have taken part in these studies sponsored by the Department of Defense since that time. The goal is to improve the standard treatment of hemorrhage and burns, which has not changed over the past 50 years. We had earlier shown that the d isomer of lactic acid, a component in Ringers Lactate, induced cardiac arrythmias in children undergoing resuscitation from burn and could induce fatal arrythmias in experimental animals [Chan L et al, Physiol Behav Sci 1994;29:383-94]. In collaboration with the Naval Blood Research Laboratory, we designed and caused to be manufactured alternatives Ringer's solutions for use in resuscitation from hemorrhage in experimental animals. Tests of these newly manufactured fluids performed at USUHS have shown that resuscitation from hemorrhage with the standard high volume Ringer's lactate protocol leads to apoptosis in the lung 24 hours after hemorrhage and that this untoward effect can be avoided if Na D-?O-hydroxybutyrate is substituted for the D,L-lactate in such fluids [Alam HB et al, J Am Col Surg 2001;193:255-63]. Lung damage after resuscitation, so called DaNang lung was a leading cause of post surgical death during the Viet Nam war. Although not admittedly, during 2003, the US fluid manufacturers changed the composition of Ringer's lactate from D,L to L lactate. This change was not done through submission of new data to FDA, but rather through the expedient of changing the definition of lactate in the US Pharmacopeia from a substance without optical rotation, to one with optical rotation. The inclusion of L-lactate in parenteral fluids, while an improvement over fluids containing D,L-lactate, is not optimum. Conditions requiring resuscitation are almost inevitably associated with hypoxia and lactic acidosis. Additional infusion of L-lactate worsens this condition and adds to the blockade of glycolysis, the major source of energy generation during periods of hypoxia as has been shown in studies by collaborators [Cross HR et al, J Mol Cell Cardiol 1995;27:1369-81]. It is our opinion that resuscitation fluids containing D-?O-hydroxybutyrate in place of L-lactate are preferable. In collaboration with the Naval Blood Laboratory, we are attempting to design and prepare such fluids made under cGMP conditions for testing in human subjects. A manuscript entitled "Severe adverse events associated with hemoglobin based oxygen carriers: role of resuscitative fluids and liquid preserved RBC" detailing the toxic effects of current resuscitation practices and how they can be avoided has been accepted for publication in the Journal of Trauma. SIGNIFICANCE OF THIS WORK TO THE PROGRAMS OF THIS INSTITUTE Ingestion of alcohol reduces the free [NAD+]/[NADH] ratio in cytoplasm of liver as does intravenous administration of l-lactate. Therefore this work has some significance to the work of this institute. However, this work is of greater significance to medical and surgical treatments in general particularly to the treatment of those engaged in combat or subject to injury from mass disasters.

摘要：九种主要无机离子在工作灌注心脏中的梯度的能量彼此接近平衡，细胞外相和细胞内相之间的电势以及ATP水解的三角肌[Masuda T等，J Biol Chiol Chem 1990; 265：20321-34]。乙醇的代谢将肝细胞的静息电潜力从-28增加到-40 mV [Veech RL等，Alch Clin Exp Res 1994; 18：1040-56]。以前，我们表明，仅更改可用的底物会改变心脏中ATP水解的三角肌，并将心脏液压工作的效率提高了28％[Kashiwaya Y等，Am J Cardiol 1997; 80：50a-64a]。由于任何形式的损伤会引起细胞离子分布的刻板印象变化，其中细胞获得了Na+，因此失去K+和膨胀，因此这些损伤的刻板印象变化可以通过对受害者受害者或燃烧的受害者的流体组成的简单变化来逆转。我们继续表明，细胞内和细胞外相之间所有九个常见的无机离子的梯度在平衡系统附近形成了一个donnan，将相位的电势与ATP水解能量有关[Veech RL等，IUBMB Life 2002; 54：241-52]。这些研究和我们较早的研究批评了现有肠胃液的组成[Veech RL，Am J Clin Nutr 1986; 44：519-51]由医学院召集的小组建议，建议对启动新的复苏流体的可行性进行研究，以启动新的复苏流体[启动新的复苏流体[natl Acadition，natl Acarts Pers，Wash DC，1999年]。 从那时起，我们就参与了国防部赞助的这些研究。目的是改善出血和燃烧的标准治疗方法，在过去的50年中，这种治疗方法没有改变。我们早些时候表明，乳酸的D异构体是乳酸林酸盐中的一种成分，诱发了烧伤复苏的儿童的心脏arrythmias，并且可能在实验动物中诱导致命的芳香性[Chan Let al，Physiol Behav Sci Sci Sci 1994; 29：383-94]。与海军血液研究实验室合作，我们设计并导致制造替代方案Ringer的解决方案用于实验动物的出血复苏。 Tests of these newly manufactured fluids performed at USUHS have shown that resuscitation from hemorrhage with the standard high volume Ringer's lactate protocol leads to apoptosis in the lung 24 hours after hemorrhage and that this untoward effect can be avoided if Na D-?O-hydroxybutyrate is substituted for the D,L-lactate in such fluids [Alam HB et al, J Am Col Surg 2001; 193：255-63]。复苏后的肺部损害，所谓的丹甘肺是越南战争期间手术后死亡的主要原因。尽管诚然，但在2003年期间，美国流体制造商将林格乳酸的组成从d，l，l乳酸更改。这种变化不是通过向FDA提交新数据来实现的，而是通过将美国药房中乳酸的定义从没有光旋转的物质变为具有光学旋转的物质的权宜之计。 在肠胃外液中包含L乳酸，而对含有D，L-乳酸的流体的改善不是最佳的。需要复苏的条件几乎不可避免地与缺氧和乳酸性酸中毒有关。 L-lactate的额外输注使这种情况恶化，并增加了糖酵解的阻断，糖酵解是缺氧期间的主要能源产生来源，如协作者所表明的[Cross Hr等，J Mol Cell Cardiol 1995; 27：1369-81]。我们认为，最好选择含有d- o-羟基丁酸的复苏液代替L-乳酸盐。与海军血液实验室合作，我们正在尝试设计和准备在CGMP条件下制成的流体，以在人类受试者中进行测试。一个名为“与基于血红蛋白的氧载体相关的严重不良事件：复苏液的作用和液体保留的RBC的作用”，详细介绍了当前复苏实践的毒性作用，以及如何避免它们在创伤杂志上被接受以发表。 这项工作对该研究所的计划的意义 摄入酒精会降低肝脏细胞质中的游离[NAD+]/[NADH]，就像静脉注射L-乳酸的给药一样。因此，这项工作对该研究所的工作具有一定的意义。但是，这项工作对医学和外科手术治疗的意义更大，特别是治疗从事战斗或遭受大规模灾难的伤害的人。