Aldosteronism: An Immunostimulatory State

描述（由申请人提供）：无论其来源如何，无症状（临床前）心室功能障碍最终发生在：循环肾素-血管紧张素-醛固酮系统的激活，伴有症状性充血性心力衰竭（CHF）和伴有血浆细胞因子升高、厌食症和消耗（临床）的全体性疾病；进行性心脏重构（病理性）。每个阶段所涉及的细胞和分子途径仍有待阐明。我们在大鼠中使用4周醛固酮（ALDO）治疗，并检查临床前（1-2周）、临床（3周）和临床病理（4周）分期。在第4周首先出现在心房和心室的血管和非血管损伤部位，在入侵的免疫细胞中发现了NADPH氧化酶表达和活性的证据，以及NFkB的激活和宿主防御编码基因的表达。与螺内酯（Spi）或n -乙酰半胱氨酸（NAC）联合治疗可预防临床和病理分期。在这里，我们假设这种促炎心脏表型是外周血单核细胞（PBMC）被氧/亚氧化应激激活并由PBMC (Mg2+)i缺乏引起的免疫刺激状态的结果。在饮食中Mg2+缺乏的wk3时，其他报告的心脏病理，同样是在wk1时PBMC激活之前，并被抗氧化剂或受体拮抗剂（L-703,606）阻止P物质，一种神经源性炎症介质。我们检测了PBMC分子表型（MP），包括所有表达的基因和蛋白质，每周从ALDO/盐处理4周的大鼠中获得。mRNA表达的变化揭示了atpase依赖基因的早期（wk1）减少和氧/亚硝化应激的进行性发展，以及持续4周以上的抗氧化防御上调。B淋巴细胞反应，包括抗体基因的上调，从一开始就出现。蛋白表达的变化确定了Mg 2+依赖性ca2 +泵和过氧化物还毒素i的早期减少。因此，PBMC的转录组和蛋白质组是应激和全身免疫刺激状态的早期标记，预示着疾病和心脏病理的发生。具体目的：1)监测体内激活的PBMC中的MP，并通过比较ALDO/盐加Mg2+在a)标准（stan）或b)缺乏（def）饮食中的反应，c) stan chow+Mg 2+补充剂，或d)单独Mg2+ def chow（无ALDO）中的反应，确定ALDO和Mg2+的作用；2)监测体内活化的PBMC的MP，通过解决Spi、NAC或L-703,606的保护作用，确定参与疾病和心脏病理出现的基因和蛋白质；3)确定在ALDO/salt/stan chow治疗反应中诱导心脏促炎表型所需的PBMC类型。这项研究可能会发现预测CHF和心脏重塑发展的临床前标志物，并可能产生治疗策略。

DESCRIPTION (provided by applicant): Irrespective of its origins, asymptomatic (preclinical) ventricular dysfunction eventuates in: activation of the circulating renin-angiotensin-aldosterone system with symptomatic congestive heart failure (CHF) and a systemic illness with elevated plasma cytokines, anorexia & wasting (clinical); and progressive cardiac remodeling (pathologic). Cellular & molecular pathways involved at each stage remain to be elucidated. We use 4wk aldosterone (ALDO) treatment in rats and examine preclinical (wks 1-2), clinical (wk3), and clinicopathologic (wk4) stages. At vascular and nonvascular sites of injury that first appear in both atria & ventricles at wk4, evidence of NADPH oxidase expression and activity is found in invading immune ceils, together with activation of NFkB and expression of genes encoding for host defenses. Clinical and pathologic stages are prevented by co-treatment with spironolactone (Spi) or N-acetylcysteine (NAC). Herein we hypothesize this proinflammatory cardiac phenotype is a result of an immunostimulatory state found in peripheral blood mononuclear cells (PBMC) activated by oxi/nitrosative stress and induced by PBMC (Mg2+)i deficiency. Cardiac pathology, reported by others at wk3 of dietary Mg2+ deficiency, is likewise preceded by PBMC activation at wkl and is prevented by antioxidant or receptor antagonist (L-703,606) to substance P, a neurogenic mediator of inflammation. We examined the PBMC molecular phenotype (MP), including all expressed genes and proteins, obtained weekly from rats treated with ALDO/salt for 4wks. Changes in mRNA expression revealed an early (wk1) reduction in ATPase-dependent genes and a progressive development of oxi/nitrosative stress, and an upregulation of antioxidant defenses sustained over 4wks. B lymphocyte responses, including upregulation of antibody genes, appear from wkl on. Changes in protein expression identified early reduction in Mg 2+-dependent Ca 2+ pump and peroxiredoxin-I. Thus, the transcriptome and proteome of PBMC serve as early markers of stress and the systemic immunostimulatory state heralding the onset of illness and cardiac pathology. Specific Aims: 1) to monitor MP in PBMC activated in vivo and to determine the role of ALDO and Mg2+ by comparing responses to ALDO/salt plus Mg 2+ in a) standard (stan) or b) deficient (def) chows, c) with stan chow+Mg 2+ supplements, or d) Mg 2+ def chow alone (no ALDO); 2) to monitor MP of PBMC activated in vivo and determine genes & proteins involved in the appearance of illness and cardiac pathology by addressing the protective effects of Spi, NAC, or L-703,606; and 3) to determine the type(s) of PBMC necessary for the induction of the proinflammatory cardiac phenotype that appears in response to ALDO/salt/stan chow treatment. This study may identify preclinical markers that predict the development of CHF and cardiac remodeling, and may produce treatment strategies.