【摘要】 目的研究HO-1对氧糖剥夺海马神经元的影响及其机制.方法将培养7d的大鼠海马神经元随机分为4组:正常培养组(C组)、正常培养+血晶素组(C+H组)、氧糖剥夺组(D组)、氧糖剥夺+血晶素组(D+H组).C组正常培养方法培养.C+H组在正常培养的神经元培养液中加入血晶素使其终浓度为10μmol/L后按正常培养24h.D组神经元进行缺糖、缺氧后复糖、复氧处理.D+H组神经元用10μmol/L血晶素处理24h后进行缺糖、缺氧后复糖、复氧处理.进行神经元纯度鉴定,细胞存活率,HO-1-mRNA表达,HO-1、Apaf-1、Caspase3蛋白表达,细胞色素C和神经元凋亡的检测.结果C+H组神经元HO-1-mRNA和HO-1表达高于C组(P<0.01),Apaf-1、Caspase3、细胞色素C、神经元存活率和神经元凋亡率变化无统计学意义(P>0.05);D组神经元HO-1-mRNA和HO-1表达高于C组(P<0.01),Apaf-1、Caspase3、细胞色素C高于C组(P<0.01),神经元存活率低于C组(P<0.01),神经元凋亡率高于C组(P<0.01);D+H组神经元HO-1-mRNA和HO-1表达高于D组(P<0.01),Apaf-1、Caspase3、细胞色素C低于D组(P<0.01),神经元存活率高于D组(P<0.01),神经元凋亡率低于D组(P<0.01).结论HO-1降低细胞色素C的释放和Apaf-1、Caspase3的表达,抑制神经元的凋亡.更多还原

【Abstract】 Objective To investigate the neuroprotective effects of hemeoxygenase-1(HO-1) on oxygen glucose deprivation(OGD) injury to primary cultured rat hippocampus neurons.Methods Newborn(24～48 h) Wistar rats were decapitated and hippocampus tissue was dissected and cut into 1mm x 1mm x1mm pieces,then digestied with 0.125% trypsin,suspended in a medium.Cells were plated at 1.0 ×105 /mL on poly-Dlysine-treated 96-well(100 μL /wel)l plates as well 6-well(2 mL /wel)l plates.Cells were used after 7 days.For oxygen glucose deprivation(OGD) experiments,cultures were washed three times in a glucose-free balanced salt solution(BSS).They were then placed in deoxygenated glucose-free medium and sealed under 95% N2 5% CO2 in an anaerobic chamber equilibrated to 37 ℃ and 100% humidity for 45 min.OGD was terminated by replacement of stored medium and by returning the cultures to a standard incubator maintained at 37 ℃ in 95% air-5% CO2.Experimental group cells were respectively carried out 10 μmol/L Heme precondition(group C+H),OGD(group D),10 μmol/L Heme precondition+OGD(group D+H),Control cells were cultured normally(group C).Compound remained present throughout the duration of the experiment until analysis 24 h later.Neuron viability and apoptosis were weasured.The expression of HO-1,Apaf-1,Caspase3 protein and HO-1-mRNA as well as cytochrome C(Cyto C) were detected.Results The expression of HO-1 protein and HO-1-mRNA increased(vs group C,P<0.01),Apaf-1,Caspase3,Cyto C,neuron viability and apoptosis had no significant change(vs group C,P >0.05) in group C+H.HO-1 protein and HO-1-mRNA decreased(vs group C,P<0.01),Apaf-1,Caspase3,Cyto C increased(vs group C,P< 0.01),neuron viability decreased and apoptosis increased(vs group C,P<0.01) in group D.HO-1 protein and HO-1-mRNA increased(vs group D,P<0.01),Apaf-1,Caspase3,Cyto C decreased(vs group D,P <0.01),neuron viability increased and apoptosis decreased(vs group D,P <0.01) in group D+H.Conclusions HO-1 protects hippocampus neurons against oxygen glucose deprivation injury through inhibiting cytochrome C releasing,and decreasing Apaf-1 and Caspase3 levels,eventually inhibiting neuron apoptosis.更多还原