摘要
目的观察血晶素(Hemin)对海马脑片氧糖剥离(OGD)模型的保护作用并探讨其机制。方法取新生8~10dSD大鼠,制备厚约400μm海马脑片,培养2周后筛取完好无特异荧光着色的脑片,分为正常培养对照组(HOTC)、缺氧缺糖组(OGD)、预加Hemin再缺氧缺糖组(Hemin+0GD)和预加血红素加氧酶-1(Heme oxygenase-1,HO-1)抑制剂锌原卟啉(Znpp)再缺氧缺糖组(Znpp+OGD)。除HOTC组外,其余各组在5%C02、95%N2的混合气体中培养1.5小时,其后恢复培养24小时。海马脑片冰冻连续冠状切片,应用免疫组织化学染色和图像分析处理技术观察海马脑片脑红蛋白(Neuroglobin,Ngb)和HO-1表达变化,并观察神经元形态学改变。结果海马脑片发生缺氧缺糖时,神经元中Ngb蛋白和HO-1蛋白表达增加,血晶素促进神经元Ngb蛋白和HO-1蛋白表达,并减轻海马脑片缺糖缺氧性损伤。Znpp抑制神经元Ngb蛋白和HO-1蛋白表达,并加重海马脑片缺糖缺氧性损伤。结论血晶素可减轻海马脑片缺糖缺氧性损伤,其机制可能通过诱导HO-1表达从而增加神经元Ngb表达发挥作用。
Objective To investigate the neuroprotective role of hemin and its effect on the expression of neuroglobin(Ngb) and heme oxygenase-1 (HO-1) in the hippocampal organotypic tissue cultures (HOTC) after combined oxygen/ glucose deprivation(OGD). Methods SD rats, 8-10-days old, were sacrificed for brain. HOTC were made and randomly divided into four groups, including HOTC, HOTC undergoing OGD (OGD), HOTC undergoing OGD following pretreatment with hemin (Hemin + OGD) and HOTC undergoing OGD following pretreatment with Znpp(Znpp+ OGD). The expression of neuroglobin and HO-1 was detected with immunohistochemical technique and computer-assisted image analysis. Results In OGD group, hippocampal organotypic tissue lost normal structure and neurons were damaged severely, and the expression of Ngb protein and HO-1 protein were significantly stronger than that in the HOTC group. Pretreatment with hemin could effectively reverse the damage of the neurons, and accompanied with upregulation of Ngb protein and HO-1 protein expression. In contrast, pretreatment with Znpp, an HO-1 inhibitor, could aggravate the damage, and accompanied with reduction of Ngb protein and HO-1 protein expression. Conclusion Hemin has protective effect on injury induced by OGD in rat hippaocampal slices, which is associated with enhanced expression of neuroglobin via the upregulation of HO-1.
出处
《西部医学》
2009年第3期357-360,共4页
Medical Journal of West China
基金
国家自然科学基金资助项目(30500170)