乙酰左旋肉碱对大鼠急性脊髓损伤后线粒体氧化应激和自噬的调节及机制研究

Acetyl-l-carnitine (ALC) is a powerful antioxidant, which can promote fat metabolism, transport energy and anti-oxidation, and have anti-inflammatory and anti-apoptosis effects. Our preliminary experiments results suggest that ALC can relieve oxidative stress reaction after spinal cord injury in rats, improve mitochondrial function and promote restoration of the neural function, but its mechanism remains controversial. A large number of studies suggested that SIRT3, a key protective factor regulating cell energy balance, plays an important role in regulating oxidative stress injury. We hypothesize that ALC could inhibit mitochondrial oxidative stress and stimulate mitochondrial autophagy by activating SIRT3/Nrf2-keap1/AER pathway , playing a nerve protective effect in spinal cord injury. Based on PC12 cell oxidative-stress model and spinal cord injury rat model, this project will start with SIRT3/Nrf2-keap1/AER pathway to study the effect of ALC on spinal cord injury, using methods of the behavioral evaluation, electron microscope, TUNEL, Western blot, Si RNA, and immunohistochemistry. The results will provide an important experimental foundation for the clinical application of ALC in the treatment of spinal cord injury.

乙酰左旋肉碱（ALC）是一种强抗氧化剂，具有促进脂肪代谢、转运能量和抗氧化、抗炎症、抗凋亡等多种生物学作用。我们的预实验结果提示ALC可以减轻大鼠急性脊髓损伤后氧化应激反应，改善组织线粒体功能促进神经功能修复，但其作用机制有待进一步探讨。大量实验提示SIRT3作为调节细胞能量平衡的关键性保护性因子，在氧化应激损伤调节中起重要作用。我们提出假说：ALC可能通过激活SIRT3/Nrf2-keap1/ARE通路抑制线粒体氧化应激和诱导线粒体自噬，对脊髓损伤发挥神经保护作用。本项目将通过细胞模型、大鼠脊髓损伤模型，采用行为学、透射电镜、TUNEL、Western blot、免疫组化和RNA干扰等方法，以SIRT3/Nrf2-keap1/ARE通路为切入点，从分子、细胞、组织以及动物整体水平等方面研究ALC对脊髓损伤修复和可能分子机制，为临床应用ALC治疗脊髓损伤提供理论和实验依据。

乙酰左旋肉碱（ALC）是一种强抗氧化剂，具有促进脂肪代谢、转运能量和抗氧化、抗炎症、抗凋亡等多种生物学作用。我们的预实验结果提示ALC可以减轻大鼠急性脊髓损伤后氧化应激反应，改善组织线粒体功能促进神经功能修复，但其作用机制有待进一步探讨。大量实验提示SIRT3作为调节细胞能量平衡的关键性保护性因子，在氧化应激损伤调节中起重要作用。我们提出假说：ALC可能通过激活SIRT3/Nrf2-keap1/ARE通路抑制线粒体氧化应激和诱导线粒体自噬，对脊髓损伤发挥神经保护作用。.本研究以H2O2制备氧化PC12细胞的损伤模型和大鼠脊髓急性打击损伤动物模型为基础，采用形态学、行为学、Western blot、TUNEL、免疫组化和RNA干扰等方法，从分子、细胞、组织以及动物水平等层次围绕以下内容进行研究：①进一步明确ALC对脊髓损伤的神经保护作用；②阐明ALC抑制线粒体氧化应激和增强线粒体自噬的作用；③探讨ALC通过调控SIRT3/Nrf2-Keap1/ARE通路发挥对脊髓损伤的神经保护作用。本课题将从ALC对脊髓损伤的药物作用新靶点SIRT3/Nrf2-Keap1/ARE通路作为研究的切入点，为ALC治疗脊髓损伤后神经修复提供理论和实验依据。