CALPAINS AND CASPASE-3 IN NEURONAL CELL DEATH

描述：（逐字摘自申请人摘要）创伤性脑损伤（TBI）是一个重要的医疗保健问题，约有50万住院患者。尽管在了解与创伤性脑损伤相关的病理机制方面取得了很大进展，但对影响创伤性脑的复杂而多样的生化、细胞和分子级联反应的了解甚少。然而，我们的实验室和其他实验室最近的进展表明，半胱氨酸蛋白酶的两个家族，钙蛋白酶和半胱天冬酶，在介导创伤后坏死和/或凋亡细胞死亡中的重要作用。最近发展的与TBE相关的各种机械和生化损伤的体外模型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calpain和caspase-3蛋白酶在各种体外坏死或凋亡模型、机械细胞损伤模型、谷氨酸兴奋性毒性和原代中隔海马细胞培养中氧/葡萄糖剥夺的相对贡献作用。重要的是，初步工作已经证明calpain和caspase-3蛋白酶可以单独或同时被激活，这取决于启动刺激的模式和/或强度。此外，这项工作证明了caspase-3激活作为检测细胞凋亡的敏感生化标志物，独立于经典定义的形态学凋亡改变。人们认为，这项研究的结果将促进对细胞损伤的各种病理反应的理解，并将为最终使用治疗干预多种中枢神经系统疾病提供良好的基础。

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.