艾片调控神经元线粒体Ca2+-MCU通路的醒神回苏机制研究

l-borneolum has effect on resuscitation and has been commonly used for stroke in clinic. Cerebral ischemic stroke is a kind of emergency and severe disease with high morbidity, high mortality and high morbidity. In our previous researches, l-borneolum was effective in hypoxia-ischemia, regulating the blood-brain barrier and cerebral protection. The effect above was thought as the basic of its "resuscitation" function and "cold" property. However, the deep mechanism remains poorly understand. The latest researches demonstrated that mitochondrial calcium uniporter (MCU) is the key protein in mediating the energy metabolism and dominantly regulating the dynamic equilibrium of energy information in mitochondria. The excessive activation of Ca2+-MCU pathway plays the key process of cerebral ischemic stroke. This project proposes a hypothesis that Ca2+-MCU signal pathway is the crucial mechanism of l-borneolum to exhibit the “cold property” – “resuscitation function”. Based on the holistic view of traditional Chinese medicine and the mitochondrial energy metabolism-apoptosis pathway mediated by MCU, we use the pharmacology and bio-thermodynamics combined with two-photon excitation fluorescence imaging and molecular simulation technology to focuse on the Ca2+-MCU signaling pathway of neuron mitochondria in ischemia model. We will systematically study the characteristics of mitochondrial energy metabolism and the biological mechanism of stability and protection of neuronal function regulated by l-Borneolum on cerebral ischemic stroke. Ultimately, this project will provide a scientific connotation and improve the “cold property” – “resuscitation function” essence of l-Borneolum.

艾片有开窍醒神回苏功效，主治中风痰厥神昏。缺血性脑卒中昏迷与神经元线粒体损伤等因素高度相关。受基金资助已从能量代谢角度初步证实艾片有抗脑缺氧缺血、调节血脑屏障，发挥脑保护作用而体现“开窍醒神”功效，并有“寒性”特征，但深层次机制尚不明确。最新研究认为：线粒体钙离子单向转运体（MCU）是介导能量代谢的关键蛋白，主导线粒体能量信息的动态平衡，Ca2+-MCU通路的过度激活在缺血性脑卒中的发病过程有重要影响。故本课题提出Ca2+-MCU信号通路是表达艾片“寒性-醒神回苏”关键机制的科学假说。拟以中医药理论思想为指导，以MCU介导的线粒体能量代谢-凋亡通路为切入点，采用药理学与生物热力学方法，结合双光子成像、分子模拟等技术，围绕线粒体Ca2+-MCU通路，系统研究艾片调控脑缺血损伤线粒体的能量代谢特征及保护神经元功能的生物学机制，为阐释艾片改善缺血性脑卒中表达“性寒-醒神回苏”的科学内涵提供参考。

艾片有开窍醒神回苏功效，主治中风痰厥神昏，临床多用于缺血性脑卒中的中药治疗，然而其功效的现代科学内涵仍有待阐释。缺血性脑卒中昏迷与神经元线粒体损伤等因素高度相关，前期研究初步证实艾片有抗脑缺氧缺血、调节血脑屏障， 发挥脑保护作用而体现“开窍醒神”功效，并有“寒性”特征，但深层次机制尚不明确。线粒体钙离子单向转运体（MCU）是介导能量代谢的关键蛋白，主导线粒体能量信息的动态平衡，Ca2+-MCU通路的过度激活在缺血性脑卒中的发病过程有重要影响。本课题融合循证医学与用药规律分析方法，证实艾片作为常用的开窍药，在治疗缺血性脑卒中方面使用频率高，推测可能与其脑保护作用密切相关，进一步以临床上包含有艾片或冰片的中成药制剂为代表，以脑卒中为代表的脑血管疾病为传统中医闭证的表征，通过Meta分析手段，明确其在防治脑卒中等方面的有效性与安全性。进一步采用生物热力学的方法，发现三种不同冰片对铜绿假单胞杆菌的热活性影响存在着显著差异，其抑制模式菌种的能量代谢强度为艾片>天然冰片>合成冰片，并明确了3种冰片对整体动物的各生理指标有抑制，是其表达“寒性”的可能特征。进一步采用药理药效学、分子生物学、代谢组学等多学科技术，揭示了艾片艾片可显著改善皮质区与海马区的脑损伤，抑制神经元的凋亡，其是通过调控Ca2+-MCU通路而稳定线粒体能量代谢-凋亡程序途径，为其表达“开窍醒神”功效的生物学机制。该课题的研究模式契合现代科学认知的艾片“性-效”关联研究新思路与新方法，以临床为导向的研究模式，为中药药性理论的现代研究提供了新途径。