CALCIUM ION DEPENDENT PHOSPHOKINASE C ISOFORMS IN ADAPTATION/INFLAMATION

适应/炎症中钙离子依赖性磷酸激酶 C 亚型

• 批准号: 6296720
• 负责人: ANIRBAN BANERJEE
• 金额: $ 21.07万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6296720
• 关键词: adenosine; apoptosis; calcium ion; cytotoxicity; heart cell; heart function; human tissue; immunocytochemistry; inflammation; isozymes; laboratory rat; macrophage; myocardial ischemia /hypoxia; neutrophil; protein kinase C; protein localization; trauma

PKC isoforms differ in structure, CA++ dependency (cPKC) and regulation by lipids, but are present in all tissues examined. The specific combination of isoforms expressed are characteristic of that cell phenotype. This suggests that these isoforms do not have overlapping functions. Although functional cardioprotection induced by preconditioning is inhibited by PKC inhibitors, it is not clear what roles the isoforms play. Several PKC- linked stimuli, including Ca++ preconditioning, mediate different patterns of isoform translocation to different myocellular compartments. Several lines of evidence suggest that each receptor stimulated, isoform-profile encodes distinct mechanisms of protection. Comparison of different post- injury outcomes indicates that stimuli conferring preconditioning against post-ischemic mechanical dysfunction do not automatically protect against cell-death. Further, different degrees of functional protection against progressively severe ischemia indicates that preconditioning stimuli differ in their potency. Significantly, outcomes such as functional protection do not appear to be simply correlated with either the number of isoform engaged or the extent of translocation, indicating the importance of spatially precise isoform translocation, for the correct duration. This leads to the hypothesis that 'PKC' mediated mechanisms of protection or inflammation may be encoded by distinct combinations of PKC isoforms in specific compartments. In this proposal we will build on previous work on cardiac functional protection to 1. evaluate the efficacy of a set of preconditioning stimuli against cardiac apoptosis and infarction that occurs after severe ischemia. Map the spatial and temporal translocation for each isoform engaged by this set of cardiac stress, receptor, and direct PKC-linked stimuli. 3. Determine the specific role of the cPKC isoforms in mediating mechanisms of cardioprotection against post-ischemic dysfunction, infarction and apoptosis. Several heart cells including resident leukocytes express cPKC isoforms. Neutrophils and macrophages are important in systemic post-traumatic inflammation and when stimulated by appropriate receptor, are known to involves PKC in their cytotoxic functions. We will therefore investigate whether PKC linked receptors also translocate unique isoform profiles in these inflammatory leukocytes, by 4. determining the spatial and temporal translocation of isoforms after prototypic inflammatory stimuli and 5. assessing the role of the cPKC isoforms in mediating the cytotoxicity of appropriately stimulated isolated human neutrophils and rat macrophages.

PKC同工型在结构，Ca ++依赖性（CPKC）方面有所不同，并通过 脂质，但存在于所有检查的组织中。特定组合 表达的同工型是该细胞表型的特征。这 表明这些同工型没有重叠的功能。虽然 PKC抑制了通过预处理引起的功能性心脏保护作用 抑制剂，尚不清楚同工型扮演什么角色。几个PKC- 链接的刺激，包括CA ++预处理，介导不同的模式 同工型转移到不同的心肌室。一些 证据线表明，每个受体均受到同工型的刺激 编码不同的保护机制。比较不同的后 伤害结果表明刺激赋予预处理 缺血后的机械功能障碍不会自动防止 细胞死亡。此外，针对不同程度的功能保护 逐渐严重的缺血表明预处理刺激 他们的效力有所不同。值得注意的是，诸如功能之类的结果 保护似乎与任何一个数量都不相关 参与的同工型或易位程度，表明重要性 在正确的持续时间内，空间精确的同工型易位。 这导致了“ PKC”介导的保护机制的假设 或炎症可以通过PKC同工型的不同组合来编码 特定隔室。在此提案中，我们将以先前的工作为基础 心脏功能保护至1。评估一组的功效 针对心脏凋亡和梗塞的预处理 在严重的缺血后发生。绘制空间和时间易位 对于这组心脏应力，受体和 直接PKC连接刺激。 3。确定CPKC的具体作用 在介导的心脏保护机制中针对缺血后的同工型 功能障碍，梗塞和凋亡。 包括常驻白细胞在内的几个心脏细胞表达CPKC同工型。 中性粒细胞和巨噬细胞在创伤后全身很重要 炎症，当被适当的受体刺激时，已知 涉及PKC的细胞毒性功能。因此，我们将调查 PKC链接的受体是否也将独特的同工型轮廓转移 这些炎症白细胞，按4。确定空间和时间 原型炎症刺激和5后同工型​​的易位。 评估CPKC同工型在介导细胞毒性中的作用 适当刺激的分离的人类嗜中性粒细胞和大鼠巨噬细胞。