CALPAINS AND CASPASE-3 IN NEURONAL CELL DEATH

描述：(逐字摘自申请者摘要)创伤性脑损伤(TBI)是一个重大的医疗保健问题，约有500,000人入院治疗。尽管在理解与脑损伤相关的病理机制方面已经取得了很大的进展，但对影响创伤脑的复杂而多样的生化、细胞和分子级联反应的了解却很少。然而，我们实验室和其他实验室的最新进展表明，两个半胱氨酸蛋白酶家族，即钙蛋白酶和半胱氨酸氨基转移酶，在介导创伤后坏死和/或凋亡细胞死亡方面发挥了重要作用。近年来，与创伤性脑损伤相关的各种机械和生化损伤的体外模型的发展为分离创伤诱导的刺激和随后的细胞反应提供了有效和强大的工具。本研究的目的是研究在原代培养的隔海马神经细胞中，钙蛋白酶和caspase-3酶在不同的体外坏死或凋亡模型以及机械细胞损伤、谷氨酸兴奋性毒性和缺氧/葡萄糖剥夺模型中的相对作用。重要的是，初步工作已经证明，根据启动刺激的模式和/或大小，Calain和caspase-3蛋白酶可以单独或同时激活。此外，本研究还证明了caspase-3的激活可作为一种敏感的生化标记物用于检测细胞凋亡，而不依赖于经典定义的细胞凋亡改变。本研究的结果将促进对细胞损伤的不同病理反应的理解，并将为最终在多种中枢神经系统疾病中使用治疗干预措施提供良好的基础。

DESCRIPTION: (Verbatim from the Applicant's Abstract) Traumatic brain injury (TBI) is a significant health care issue with approximately 500,000 hospital admissions. Although there has been much progress in understanding he pathological mechanisms associated with TBI, knowledge of the complex and varied biochemical, cellular and molecular cascades affecting the traumatized brain are poorly understood. However, recent advances by our laboratory and others have demonstrated important roles for two families of cysteine proteases, calpains and caspases, in mediating necrotic and/or apoptotic cell death after trauma. Recent development of in vitro models of various components of mechanical and biochemical insults relevant to TBE have provided efficient and powerful tools for dissociation of trauma induced stimuli and subsequent cellular responses. The aim of this proposal is to examine the relative contributory roles of calpains and caspase-3 proteases in various in vitro models of necrosis or apoptosis and in models of mechanical cell injury, glutamate excitotoxicity , and oxygen/glucose deprivation in primary septo-hippocampal cell cultures. Importantly, preliminary work has demonstrated that calpains and caspase-3 proteases can be individually or concurrently activated depending on the mode and/or magnitude of the initiating stimulus. In addition, this work demonstrates the use of caspase-3 activation as a sensitive biochemical marker for detection of apoptosis that is independent of classically defined morphological apoptotic alterations It is thought that result of this research will advance an understanding of the varied pathological responses to cellular insult, and will provide a sound foundation for eventual use of therapeutic interventions in numerous disorders of the central nervous system.