CELL NA IN VSM PROTEIN PRODUCTION

VSM 蛋白质生产中的细胞 NA

• 批准号: 3347028
• 负责人: Julius Allen
• 金额: $ 19.01万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-01-01 至 1992-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3347028
• 关键词: aldosterone; amiloride; angiotensin II; cell morphology; cyclic AMP; dogs; gel electrophoresis; hypertension; intermediate filaments; membrane permeability; membrane proteins; muscle contraction; muscle proteins; nucleic acid inhibitor; protein biosynthesis; tissue /cell culture; vascular smooth muscle; veratrum alkaloid

The basic underlying hypothesis of this proposal is that (Na+)i plays a role in regulating the amount of contractile proteins and the number of Na+ pump sites in cultured vascular smooth muscle cells. We will grow both arterial and venous smooth muscle cells in culture and quantitate contractile protein content (actin and myosin) by gel electrophoresis, Na+ pump site number by H3-ouabain binding and Na+ pump turnover rate by ouabain sensitive Rb86 uptake. After control characteristics have been established for these protein complexes, the cells will be grown under 4 specific conditions which will alter (Na+)i by significantly varied mechanisms: 1) O K+ or ouabain, which will inhibit the Na+ pump and increase (Na+)i; 2) monensin, aldosterone, angiotensin II and veratridine, which will increase (Na+)i and stimulate the pump; 3) amiloride which will decrease (Na+)i and inhibit the pump; and 4) cyclical stretch. The contractile protein content and Na+ pump site number will then be assessed after the appropriate growth period. In addition to these specific parameters, important additional parameters will be assessed to characterize the affect of these altered conditions. These will include determining the cell content of cAMP, DNA, Na+, K+, Ca++, the measurement of cell volume, intermediate filament content and H3-thymidine incorporation into DNA and morphological characterization (scanning and TEM). If an increase in contractile protein content or Na+ pump site number occurs, the role of Ca++ and amino acids in this process will be assessed by determining the amont of Na+i-Ca++o exchange and H3-alpha aminoisobutyric acid uptake. In addition, the effect of protein synthesis inhibitors (anisomycin, cyclohexamide) will be determined. Assessment of the amount of contractile protein and the number of Na+ pump sites under these systematically controlled conditions will lead to a better understanding concerning the role of the (Na+)i in regulating the amount of these proteins. Because of the similarity between these growth conditions and certain models of hypertension which involve increased (Na+)i, increased strain, and Na+ pump alterations, this work may lead to a hypothesis regarding the etiology of this disease.

该提议的基本基本假设是（Na+）i扮演 在调节收缩蛋白量和Na+的数量中的作用 培养的血管平滑肌细胞中的泵部位。 我们将成长 培养和定量的动脉和静脉平滑肌细胞 凝胶电泳的收缩蛋白含量（肌动蛋白和肌球蛋白），Na+ H3- ouabain结合和Na+泵的周转率通过泵位点编号 Ouabain敏感RB86吸收。 在控制特征之后 为这些蛋白质复合物建立，细胞将在4下生长 特定条件会改变（Na+）i的明显变化 机制：1）o k+或ouabain，将抑制Na+泵和 增加（Na+）i； 2）Monensin，醛固酮，血管紧张素II和Veratridine，， 会增加（Na+）i并刺激泵； 3）Amiloride将 减少（Na+）i并抑制泵； 4）周期性伸展。 这 然后将评估收缩蛋白含量和NA+泵送站点编号 在适当的生长期之后。 除了这些特定 参数，重要的其他参数将被评估为 表征这些改变条件的影响。 这些将包括 确定CAMP，DNA，Na+，K+，Ca ++的细胞含量，测量 细胞体积，中间细丝含量和H3-胸苷 纳入DNA和形态表征（扫描和 tem）。 如果收缩蛋白含量增加或Na+泵位点增加 发生数字，Ca ++和氨基酸在此过程中的作用将是 通过确定Na+I-Ca ++ o交换和H3-Alpha的即将进行评估 氨基异丁酸的摄取。 另外，蛋白质合成的作用 将确定抑制剂（砷霉素，环己酰胺）。 评估 收缩蛋白的量和Na+泵位点的数量 这些系统控制的条件将导致更好 了解（Na+）I在调节量中的作用 这些蛋白质。 由于这些增长条件之间的相似性 以及某些涉及增加（Na+）I的高血压模型 应变增加和Na+泵的改变，这项工作可能导致 关于该疾病的病因的假设。