REGULATION OF ANF AND BNP IN CARDIAC HYPERTROPHY

ANF 和 BNP 在心肌肥厚中的调节

• 批准号: 2211298
• 负责人: MARGOT CLAIRE LAPOINTE
• 金额: $ 6.37万
• 依托单位: HENRY FORD HEALTH SYSTEM
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2211298
• 关键词: atrial natriuretic peptide; chemical structure function; gel mobility shift assay; gene expression; genetic promoter element; genetic transcription; northern blottings; polymerase chain reaction; posttranscriptional RNA processing; reporter genes; saluretic; tissue /cell culture; transcription factor; transfection; vasoconstrictors; ventricular hypertrophy

Atrial natriuretic factor (ANF) and brain natriuretic peptide (BNP) have vasodepressor effects, while in the kidney they have potent natriuretic and diuretic activity. Although they have similar biological activity, each hormone has distinct features, most importantly differences in DNA regulatory sequences involved in gene expression. In adult hearts, the gene for ANF is preferentially expressed in the atria, and ventricular expression is less than 5% of the whole heart. In contrast, ventricular BNP mRNA represents greater than 70% of whole heart levels. ANF and BNP mRNA are augmented in the ventricle under various conditions of volume and pressure overload, including hypertension and heart failure. Given that the BNP gene is expressed primarily in the ventricles while the ratio of BNP to ANF is different in atria and ventricles there must be clear differences in the way ANF and BNP synthesis are regulated. The objective of this proposal is to define transcriptional and post-transcriptional mechanisms involved in differential regulation of the ANF and BNP genes during cardiac hypertrophy using 2 in vitro models of hypertrophy: adult feline cardiocytes stimulated to beat with isoproterenol and phenylephrine- or PMA-stimulated neonatal ventricular cardiocytes. Specifically, the hypotheses to be tested are that: 1) there are differences in the rates of transcription of the ANF and BNP genes, and cis-acting regulatory elements in the 5' flanking sequences (fS) of the BNP gene are distinct from those characterized for the ANF gene in neonatal cardiocytes; 2) during cardia hypertrophy, re-expression of the ANF and BNP genes involves cis-acting regulatory regions distinct from those involved in normal tissue-specific gene expression; 3) different trans-acting regulatory proteins affect ANF and BNP gene expression in normal vs. hypertrophied cardiac myocytes; and 4) BNP mRNA stability is regulated differently in normal and hypertrophied ventricles. These studies will employ cell culture models of hypertrophy, Northern blot of mRNA, transfection of chimeric promoter-reporter gene constructions, enzyme assays (luciferase and beta-galactosidase), polymerase chain reaction (PCR), DNA- and RNA-protein gel shift, and in vivo analysis of regulator elements (in vivo injection of DNA). Thus they will provide a detailed analysis of differences in ANF and BNP gene regulation and help us understand the relevance of each hormone in disease, such as hypertension and hypertrophy.

心房亚催化因子（ANF）和脑脂肪酸肽（BNP）具有 血管抑制剂效应，而在肾脏中，它们具有有效的纳地尿素 和利尿活动。尽管它们具有相似的生物学活性，但 每种激素具有不同的特征，最重要的是DNA的差异 与基因表达有关的调节序列。在成人心中 ANF的基因在心房中优先表达 表达不到整个心脏的5％。相反，心室 BNP mRNA占整个心脏水平的70％以上。 ANF和BNP 在各种体积条件下，心室中的mRNA被增强 压力超负荷，包括高血压和心力衰竭。鉴于 BNP基因主要在心室中表达 BNP到ANF的心房有所不同，心室必须有清晰的 ANF和BNP合成的方式的差异受到调节。目标 该建议的内容是定义转录和转录后 涉及ANF和BNP基因差异调节的机制 在心脏肥大期间使用2种体外肥大模型：成人 猫的心脏细胞刺激以异丙肾上腺素和 苯肾上腺素或PMA刺激的新生儿心室心细胞。 具体而言，要测试的假设是：1） ANF和BNP基因转录率的差异，以及 在5'侧翼序列（FS）中的顺式作用调节元件 BNP基因与在ANF基因中的特征的基因不同 新生儿心细胞； 2）在心脏肥大期间，重新表达 ANF和BNP基因涉及从不同的调节区域 参与正常组织特异性基因表达的人； 3）不同 跨作用调节蛋白影响ANF和BNP基因表达 正常与肥大心肌细胞； 4）BNP mRNA稳定性是 在正常和肥厚的心室中受到不同的调节。这些 研究将采用肥大的细胞培养模型，北印迹 mRNA，转染嵌合启动子 - 重生基因构建体， 酶测定（荧光素酶和β-半乳糖苷酶），聚合酶链 反应（PCR），DNA和RNA-蛋白凝胶移位以及体内分析 调节元件（体内注射DNA）。因此，他们将提供 详细分析ANF和BNP基因调节的差异并有助于 我们了解每种激素在疾病中的相关性，例如 高血压和肥大。