PGRN对脑缺血后的神经再生和神经保护作用及机制研究

Neurogenesis and neuroprotection play important roles in the recovery of brain structure and function after cerebral ischemia. Our previous data showed that progranulin （PGRN）was upregulated in blood of ischemic stroke patients and in brain tissue of ischemic rats. PGRN has been reported to be a mitogen, neurotropic and anti-inflammatory protein. Thus, we hypothesized that PGRN may involve in the neurogenesis and neuroprotection after cerebral ischemia. In this project, hypoxic response element (HRE) was used to construct lentiviral vector expressing rat PGRN cDNA and shRNA-PGRN against rat PGRN, and the developed lentiviral vectors were applied to neural stem cells(NSC)/neural progenitor cells (NPC) or the brain of the rats. Then the effect of PGRN on NPC proliferation and differentiation, on nurite outgrowth and synaptic composition, and on neural cell survival, will be investigated in hypoxic cultured NSC/NPC (in vitro) and pMCAO model (in vivo) by using in situ hybridization, immunofluorescence, confocal laser, and behavioral observation et al. The roles of PGRN in MAPK, PI3K，casepase and TNF pathways will be further studied. The results will help to understand the roles and mechanism of PGRN in neurogenesis and neuroprtection after cerebral ischemia, and may provide a new therapeutic target for ischemic stroke and other neurodegenerative diseases.

促进脑缺血后的神经再生和神经保护作用对脑结构和功能的恢复至关重要。我们前期观察到PGRN在脑缺血病人血液及缺血大鼠脑组织表达上调。PGRN具有促进细胞有丝分裂、神经营养和抑制炎性反应等作用，可能参与脑缺血后的神经再生和神经保护过程。本研究拟用缺氧反应元件HRE构建表达PGRN cDNA和 针对PGRN 的shRNA慢病毒载体以调控PGRN的表达，然后利用免疫荧光、实时定量PCR、Western blot、共聚焦显微镜和行为学观察等技术，在细胞及动物模型水平，观察缺血缺氧性损伤后PGRN对神经干细胞/前体细胞增殖分化、神经元突起和突触生长、神经保护以及炎性反应等的影响，并进一步研究脑缺血后PGRN对下游MAPK、PI3K、casepase和TNF通路的调控作用，探讨PGRN对大鼠脑缺血诱导的神经再生和神经保护作用及机制，为以PGRN为靶点对脑缺血及相关疾病的基因治疗提供理论基础和实验依据。

促进脑缺血后的神经再生和神经保护作用对脑结构和功能的恢复至关重要。我们前期观察到PGRN在脑缺血病人血液及缺血大鼠脑组织表达上调。PGRN具有促进细胞有丝分裂、神经营养和抑制炎性反应等作用，可能参与脑缺血后的神经再生和神经保护过程。本研究通过外源性PGRN进行在体和体外干预，然后利用免疫荧光、实时定量PCR、Western blot、共聚焦显微镜和行为学观察等技术，在细胞及动物模型水平，观察缺血缺氧性损伤后PGRN对神经干细胞/前体细胞增殖分化、神经元突起和突触生长、神经保护以及炎性反应等的影响，并进一步研究脑缺血后PGRN对下游MAPK、PI3K、casepase和TNF通路的调控作用，探讨了PGRN对大鼠脑缺血诱导的神经再生和神经保护作用及机制，为以PGRN为靶点对脑缺血及相关疾病的基因治疗提供理论基础和实验依据。