CALCIUM ION DEPENDENT PHOSPHOKINASE C ISOFORMS IN ADAPTATION/INFLAMATION

PKC亚型在结构、CA++依赖性(CPKC)和受 脂类，但在所有被检查的组织中都存在。具体的组合 所表达的亚型是该细胞表型的特征。这 这表明这些异构体没有重叠的功能。虽然 蛋白激酶C抑制预适应诱导的功能性心脏保护 作为抑制剂，目前还不清楚这些异构体扮演着什么角色。几个PKC- 相关刺激，包括钙离子预适应，调节不同的模式 异构体易位到不同的肌细胞室。几个 一系列证据表明，每个受体都受到刺激，异构体 编码不同的保护机制。不同岗位的比较- 损伤结果表明，诱导预适应的刺激对 缺血后机械功能障碍不能自动预防 细胞死亡。此外，不同程度的功能保护对 渐进性严重缺血表明预适应刺激 它们的效力是不同的。值得注意的是，结果如功能性 保护似乎并不是简单地与 异构体参与或易位的程度，表明重要性 在正确的持续时间内，空间上精确的亚型易位。 这导致了一种假设，即‘PKC’介导的保护机制 或者炎症可能由不同的PKC亚型组合编码 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.评价一套心功能保护剂的疗效 抗心肌细胞凋亡和梗死的预适应刺激 发生在严重的缺血之后。绘制空间和时间移位图 对于这一组心脏应激、受体和 与PKC相关的直接刺激。3.确定中心的具体作用 心肌缺血后保护机制中的同工酶 功能障碍、脑梗塞和细胞凋亡。 包括常驻白细胞在内的一些心脏细胞表达cPKC亚型。 中性粒细胞和巨噬细胞在全身创伤后中的重要作用 炎症和当被适当的受体刺激时，已知 参与PKC的细胞毒作用。因此，我们将调查 PKC连接的受体是否也将独特的异构体转位到 这些炎性白细胞，由4.决定空间和时间 典型炎症刺激后异构体的移位和5。 评估cPKC亚型在介导细胞毒作用中的作用 适当刺激分离的人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.