CALCIUM ION DEPENDENT PHOSPHOKINASE C ISOFORMS IN ADAPTATION/INFLAMATION

PKC亚型在结构、CA++依赖性（cPKC）和调节方面不同， 脂质，但存在于检查的所有组织中。的特定组合 是该细胞表型的特征。这 表明这些同种型没有重叠的功能。虽然 PKC抑制预处理诱导的功能性心肌保护作用 抑制剂，目前尚不清楚这些亚型发挥什么作用。几个PKC- 连锁刺激，包括Ca++预处理，介导不同的模式 同种型易位到不同的肌细胞区室。几 一系列证据表明，每种受体刺激，异构体分布 编码不同的保护机制。不同岗位的比较 损伤结果表明，刺激赋予预处理， 缺血后机械功能障碍不能自动保护 细胞死亡此外，不同程度的功能保护， 进行性严重缺血表明预处理刺激 它们的效力不同。重要的是，功能性等成果 保护似乎并不简单地与数量相关， 异构体参与或易位的程度，表明重要性 在正确的时间内进行空间上精确的异构体移位。 这导致了“PKC”介导的保护机制的假设， 或炎症可能是由PKC亚型的不同组合编码的， 特定的隔间。在本提案中，我们将在以前工作的基础上， 心功能保护1.评估一组 预处理刺激对抗心肌细胞凋亡和梗死， 发生在严重缺血后。绘制时空易位图 对于由这组心脏应激、受体和 直接PKC相关刺激。3.确定cPKC的具体作用 异构体介导缺血后心脏保护机制 功能障碍、梗死和凋亡。 一些心脏细胞，包括常驻白细胞表达cPKC亚型。 中性粒细胞和巨噬细胞在全身创伤后 已知当炎症被适当受体刺激时， 在它们的细胞毒性功能中涉及PKC。因此我们将调查 PKC连接的受体是否也在细胞中转位独特的同种型谱， 这些炎性白细胞，由4.确定空间和时间 原型炎性刺激后同种型的易位和5. 评估cPKC亚型在介导细胞毒性中的作用， 适当刺激分离的人嗜中性粒细胞和大鼠巨噬细胞。

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.