吸入麻醉药致发育期神经元毒性的线粒体动力学机制及干预研究

q吸入麻醉药在临床应用广泛，它对患儿发育期神经系统的影响受到广泛的关注。申请者前期基础研究发现，吸入麻醉药异氟烷可致发育期神经元内钙稳态失衡，引起神经元凋亡等退行性病变。神经元内钙稳态失衡可导致线粒体动力学异常，提示线粒体动力学异常可能参与了吸入麻醉药致发育期神经元毒性。本项目拟选择胎鼠或者新生大鼠，从线粒体、细胞、整体动物水平，探讨吸入麻醉药作用下发育期神经元线粒体和细胞内Ca2+的变化、线粒体动力学的改变状况、线粒体动力学调节蛋白表达变化并确定参与这种变化的信号通路；结合逆转录病毒技术，筛选出可能的干预靶点，揭示线粒体动力学异常在吸入麻醉药致发育期神经元毒性中的作用，为预防和治疗这种神经毒性提供崭新的思路。

Inhalational anesthetics are commonly administered in the clinical anesthesia and widespread concern was placed on their influence on the developmental neural system. The previous animal researches have shown that isoflurane induced developmental apoptotic neurodegeneration by disruption of the cytoplasmic calcium homeostasis. The dysregulation of calcium could lead to the imbalance of mitochondrial dynamics, which implies that abnormality of mitochondrial dynamics might be involved in the developmental neurotoxicity induced by inhalational anesthetics. The isolated mitochondria，primary-cultured neurons of hippocampi and neonatal rats are selected for this experimental programs. For mitochondrial and cytoplasmic calcium measurements, 5-day-cultured rat hippocampal neurons are preloaded with Rhod-2 and fluo-4AM,respective indicator of mitochondrial and cytoplasmic calcium, and then are imaged under laser scanning confocal microscope. To assay the dynamic change of mitochondrial morphology, the neurons are stained with MitoTracker after exposed to inhalational anesthetics.The mRNA and protein are assessed by real-time PCR and Western blot respectively after the neurons and animals are treated with inhalational anesthetics. Also, pathways of cell signaling transduction relevant to the imbalance of mitochondrial dynamics induced by inhalational anesthetics are investigated. Combined with the anti-virus technology, the probable targets for prevention and treatment of the neurodegeneration are selected. All the designed experiments are conducted to elucidate the role of mitochondrial dynamics disruption in the developmental neurotoxicity triggered by inhalational anesthetics and provide a new prospective for prophylaxis and medication of the neurotoxitiy.