臂旁核区域损伤致长时程“昏迷样”动物模型建立及神经机制研究

The establishment of animal model of chronic disorders of consciousness is the focus and difficulty of modern neuroscience consciousness research. The existing acute coma models can’t reflect the pathological process of chronic disorders of consciousness，so it is difficult to explore the mechanism of consciousness and some clinical interventions. The parabrachial nucleus, located in the upper part of the pontine tegmental, has been shown to be involved in the neural regulation of consciousness. Our preliminary work found that local hemorrhage caused by injection of collagenase IV into the parabrachial nucleus of rats could lead to disorder of consciousness. It is expected to establish long-time "coma-like" model in rats under a specific concentration and dose of collagenase IV. Combined with the early study, we will use animal EEG signal acquisition to record the sleep cycle and structure during the process of disorder of consciousness, as well as reactivate the neurons in the parabrachial nucleus with chemogenetic technology to restore a certain extent of consciousness. Immunofluorescence will be used to confirm the neural pathway of chronic disorders of consciousness, projecting to the basal forebrain, thalamus, hypothalamus and cortex downstream from the parabrachial nucleus region. The changes of metabolic molecules in related nuclei will be further explained by means of metabonomics, expounding the neural mechanism of chronic disorders of consciousness. The research results will lay the foundation for the study of consciousness, the intervention of chronic disorders of consciousness and the search for effective therapeutic targets.

慢性意识障碍动物模型的建立是现代神经科学意识研究的重点和难点。现有模型因昏迷时间太短而不能真正反映其真实世界，致使意识及意识障碍发生机制的研究以及许多临床干预手段难以开展。臂旁核区域位于脑桥被盖上部，已被证明参与意识的神经调控。本项目组前期研究发现在大鼠臂旁核区域注射胶原酶IV造成局部出血能使其出现意识障碍状态，用特定浓度和剂量的胶原酶IV有望建立大鼠长时程“昏迷样”模型。结合前期基础，我们将应用动物脑电EEG信号采集记录其意识障碍过程中的睡眠周期和结构，化学遗传技术重新激活臂旁核区域神经元使大鼠的意识得以一定程度恢复。利用免疫荧光技术证明慢性意识障碍的神经通路是以臂旁核区域为源点投射至其下游的基底前脑、丘脑、下丘脑至皮层，并结合代谢组学技术进一步说明相关核团代谢分子改变，初步阐述慢性意识障碍的神经机制。最终为意识的研究及慢性意识障碍的干预及神经调控寻找有效治疗靶点奠定基础。

慢性意识障碍动物模型的建立是现代神经科学意识研究的重点和难点。臂旁核区域位于脑桥被盖上部，已被证明参与意识的神经调控。本项目通过在大鼠臂旁核区域注射内皮素造成局部缺血建立了“昏迷样”模型，且建立了“大鼠昏迷恢复量表”，定量评估术后大鼠从无意识反应状态到有意识反应状态的神经状态改变。利用免疫荧光等技术证明慢性意识障碍的神经通路是以臂旁核区域为源点投射至其下游的基底前脑、丘脑、下丘脑至皮层，并结合MALDI-TOF MS质谱成像、单细胞测序、代谢组学技术进一步说明相关核团代谢分子改变，初步阐述慢性意识障碍的神经机制。最终为意识的研究及意识障碍的干预及神经调控寻找有效治疗靶点奠定基础。