摘要
目的观察氧-糖剥夺致皮层神经元一氧化氮(NO)的变化及对Caspase-3的影响。方法体外原代培养神经元,选取培养7d的神经元随机分为3组:正常对照组(A组),氧-糖剥夺损伤组(B组),细胞损伤+L-N-硝基精氨酸甲脂(L-NAME)干预组(C组)。采用硝酸还原法检测各组培养液中NO的含量,Western blot检测Caspase-3、神经元特异性烯醇化酶(neuron-specific enolase,NSE)蛋白表达,半定量RT-PCR检测Caspase-3mRNA表达。结果 B组培养液中NO的含量为(1.77±0.17)μmoL/L,与A组(0.93±0.15)μmoL/L和C组(0.99±0.10)μmol/L比较差异有统计学意义(均P<0.05);同时,B组的Caspase-3蛋白和mRNA表达明显高于A组和C组,B组的NSE蛋白表达明显低于A组和C组。结论氧-糖剥夺可导致皮层神经元NO产生过量,并造成神经元的损伤;L-NAME能减少NO的产生,从而减弱氧-糖剥夺导致皮层神经元损伤。
Objective To study the effect of oxygen-glucose deprivation on the content of nitric oxide （NO） and expression of Caspase-3 in cortical neurons of rat.Methods The cortical neurons were isolated from rats and cultured for 7 days.They were randomly divided into three groups：Group A,the control group ; group B,the oxygen-glucose deprivation group ; group C,the cell injury plus L-NAME group.The NO content was detected by nitroreduction assay.Expression levels of neuron-specific enolase （NSE） and Caspase-3 proteins were measured by Western blot.The mRNA level of Caspase-3 was detected by reverse transcription polymerase chain reaction （RT-PCR）.Results The NO content was significantly higher in group B than in group A [（1.77±0.17） μmoL/L vs.（0.93±0.15） μmoL/L] and group C [（1.77±0.17） μmoL/L vs.（0.99±0.10） μmol/L] （P 〈0.05）.The protein and mRNA levels of Caspase-3 were lower in groups A and C than in group B （P〈0.05）.The NSE protein level was significantly decreased in group B when compared with that in groups A and C （P〈0.05）.Conclusion Oxygenglucose deprivation may lead to excessive production of NO in cortial neurons,thereby causing neuron injury.L-NAME can ameliorate the cortical neuron injury caused by oxygen-glucose deprivation by reducing the production of NO.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2014年第5期550-553,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.81360186)
新疆生产建设兵团博士基金资助项目(No.2011BB016)
