促红细胞生成素对神经血管单元缺血缺氧损伤后的血管新生和神经保护作用分析-神经病学专业论文.docxVIP

Part II Benificial effects and possible mechanisms of recombinant human erythropoietin posttreatment on neurovascular unit in OGD injured model: an in vitro study Objective To study the benificial effects and possible mechanisms of recombinant human erythropoietin (rH-EPO) on neurovascular unit (NVU) after oxygen glucose (OGD) in vitro. Methods Three cell lines were cocultured to construct an NVU model in vitro. Transmission electron micronmicroscopy, biochemical method, fluorescence method et al.were used to test and validate it’s structure and function. The model was subjected to OGD and stimulated by rH-EPO just at the beginning of the reoxygenation procedure. The apoptotic EA.hy 926 and SH-SY5Y cells stained with Annexin V-FITC/PI were mesured, and the oxidative (MDA)and antioxidative(SOD) indicators were detected too.The expression of inducible nitric oxide synthase(iNOS) protein and caspase-3 mRNA in the cytoplasm of SH-SY5Y were also detected. Results After coculturing for 7 days, the model presented the specific structure and function of BBB, e g. positive γ-GT, tight junction and so on. OGD induced a significant increase in the apoptotic rate of EA.hy926 and SH-SY5Y, and the expression of iNOS and caspase-3 in the SH-SY5Y cell. Meanwhile, the oxidative and antioxidative system was in state of imbalance. However, rH-EPO could reverse the above mentioned damage. Conclution On the whole, the NVU model possess the BBB specific structure and function; posttreatment of NVU with rH-EPO at the beginning of reperfusion produced a significant reduction in the increased injury which caused by OGD.The possible mechanisms may include decreasing cell apoptosis , anti-free radical damage and regulating the expression of iNOS and caspase-3 and so on. 【Key words】 erythropoietin；oxygen glucose deprivation； neurovascular unit ； coculture； blood brain barrier ；posttreatment ; protection 英文缩略词及中文对照 EPO rH-EPO EPOR PBS FBS ELISA PCR Real