CHARACTERIZING NOVEL STRESS-RELATED PROTEINS IN MYOCYTES

表征心肌细胞中与压力相关的新型蛋白质

• 批准号: 6490646
• 负责人: PATRICE M CONNELL
• 金额: $ 3.58万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-01-01 至 2002-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6490646
• 关键词: JUN kinase; antisense nucleic acid; apoptosis; cardiac myocytes; cell differentiation; cell growth regulation; cell proliferation; complementary DNA; cysteine endopeptidases; cytoprotection; gene expression; genome; molecular chaperones; muscle cells; northern blottings; nuclear factor kappa beta; oxidative stress; protein binding; protein degradation; protein structure function; stress proteins; striated muscles; tissue /cell culture; transfection; western blottings

Cardiovascular disease is the sum total of a series of molecular and cellular responses to stressful stimuli. An increase in expression of stress proteins such as heat shock protein 70 (Hsp70) is a common cellular response to oxidative stress, cytokine release and ischemia-reperfusion. Its expression is correlated with the protection of cardiomyocytes and the whole heart in the event of these environmental insults. The mechanisms by which Hsp70 exerts its protective effects are as yet undetermined, but may involve its role as a molecular chaperone and its association with protein degradation pathways. Hsp70's involvement in these processes is regulated both transcriptionally and by co-chaperones. This proposal involves a recently described negative regulator of Hsp70/Hsc70 activity, carboxyl terminus Hsc70-interacting protein (CHIP). The objective of this proposal is to determine the effect of CHIP on cellular pathways/processes influenced by Hsp70 expression with regards to CHIP's ability to alter Hsp70's activity. CHIP expression will be manipulated in cardiomyoblasts (H9c2) and myocytes (C2C12) using a retroviral-mediated gene transfer technique. Overexpression of CHIP will be the focus of investigations regarding the involvement of CHIP in cell pathways known to be affected by Hsp70 and of particular relevance to cardiovascular disease. Cells will be treated with known inducers of oxidative stress and apoptosis. Production of cytokines, NF-kB activity, and specific activators (SAPK/JNK) and effectors (caspase-activated enzymes) in the apoptotic cascade will be analyzed. This work will determine if CHIP is involved in mediating Hsp70's protection in response to stressful stimuli which are known inducers of atherogenesis and cardiovascular disease.

心血管疾病是一系列对压力刺激的分子和细胞反应的总和。 诸如热休克蛋白70（HSP70）等应激蛋白表达的增加是对氧化应激，细胞因子释放和缺血 - 再灌注的常见反应。 在这些环境侮辱的情况下，它的表达与心肌细胞的保护和整个心脏有关。 HSP70发挥其保护作用的机制尚未确定，但可能涉及其作为分子伴侣的作用及其与蛋白质降解途径的关联。 HSP70参与这些过程都受到转录和副伴侣的调节。 该建议涉及最近描述的HSP70/HSC70活性的负调节剂，羧基末端HSC70相互作用蛋白（CHIP）。 该建议的目的是确定芯片对芯片改变HSP70活性的能力，受HSP70表达影响的细胞途径/过程的影响。使用逆转录病毒介导的基因转移技术，将在心肌细胞（H9C2）和心肌细胞（C2C12）中操纵芯片表达。 芯片的过表达将是有关CHIP参与已知受HSP70影响的细胞途径的重点，与心血管疾病特别相关。 细胞将用已知的氧化应激和凋亡诱导剂处理。将分析细胞因子，NF-KB活性和特定活化剂（SAPK/JNK）以及凋亡级联反应中效应子（caspase激活酶）的产生。 这项工作将确定芯片是否参与介导HSP70的保护，以应对压力性刺激，这是已知的动脉粥样硬化和心血管疾病的诱导者。