5-LO表达对铝盐致海马神经元损伤的影响  

Effect of 5-LO expression on primarily cultured rat hippocampal neuron damage induced by aluminum

作　　者：王健峰[1] 杨俊卿[1] 黄砚[1] 谢灵瑶[1] 郭远新[1] 雷文娟[1] 

机构地区：[1]重庆医科大学药理学教研室,重庆400016

出　　处：《中国药理学通报》2013年第5期664-669,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 30672211;81070972)

摘　　要：目的观察5-LO表达对铝盐致原代培养海马神经元损伤的影响。方法将5-LO过表达重组体腺病毒(Ad5-LO)和5-LO RNAi重组体腺病毒(Ad5-LO-RNAi)转染原代培养7 d的海马神经元,实验随机分为7组:空白对照组(NaCl 200μmol.L-1)、空载腺病毒组(MOI=100)、5-LO过表达腺病毒组(MOI=100)、5-LO RNAi腺病毒组(MOI=100)、空载腺病毒(MOI=100)+铝盐负荷组(AlCl3200μmol.L-1)、5-LO过表达腺病毒(MOI=100)+铝盐负荷组(AlCl3200μmol.L-1)、5-LO RNAi腺病毒(MOI=100)+铝盐负荷组(AlCl3200μmol.L-1)。采用荧光显微镜观察海马神经元形态变化,以RT-PCR和Western blot方法分别检测神经元5-LO mRNA和蛋白表达,以MTT测定和LDH漏出率检测神经元存活情况,以生化方法测定海马神经元SOD活性和MDA含量变化。结果空载腺病毒+铝盐负荷组神经元数目明显减少,胞体萎缩,突起减少变短,5-LO mRNA和蛋白表达增多,MTT值降低,LDH漏出率增多,SOD活性降低,MDA含量增加。与空载腺病毒+铝盐负荷组比较,5-LO过表达腺病毒处理使铝盐负荷神经元上述指标的变化更加明显;5-LO RNAi腺病毒处理能明显阻遏铝盐负荷组海马神经元上述指标的变化。结论 5-LO过表达可增加大鼠海马神经元对铝盐损伤的易感性,而抑制5-LO表达能够减轻铝盐对大鼠海马神经元的损伤。Aim To investigate the effect of 5-LO expression on primarily cultured rat hippocampal neurons damage induced by aluminum. Methods Rat hippocampal neurons were primarily cultured for 7 days and treated by recombinant adenovirus expressing 5-LO （Ad5-LO） and Ad5-LO-RNAi. Primarily cultured neuron were divided into 7 groups, i.e. control group （200 μmol.L-1 NaCl）, adenovirus-treated group （ MOI = 100） , Ad5-LO-treated group （ MOI = 100 ）, Ad5-LO- RNAi-treated group （ MOI = 100 ） , adenovirus （ MOI = 100） ＋ aluminum（ 200 μmol.L-1 AlCl3）-treated group, Ad5-LO （ MOI = 100） ＋ aluminum （ 200 μmol.L-11 A1C13 ）-treated group and Ad5-LO-RNAi （MOI = 100） ＋ aluminum （200 p^mol. L-1 A1C13）-treated group. The hippocampal neuron damage was determined by observation of the change of neuronal morphology and detection of SOD activity and MDA content in hippocampal neurons. The levels of 5-LO proteins and 5-LO mRNA expression were also measured. Results As compared with adenovirus-treated group, the neurons were injured remarkably ,the optical density of MTF and activity of SOD were decreased, the leakage of LDH and content of MDA and expression of 5-LO mRNA and proteins were increased in adenovirus ＋ aluminum-treated group. As compared with adenovirus ＋ aluminum group, these changes were more obvious in Ad5-LO ＋ aluminum-treated group. The treatment of Ad5-LO-RNAi could significantly blunt the changes by aluminum. Conclusions The overexpression of 5-LO increases the sensibility of hippocampal neuron to aluminum injury and the inhibition of 5-LO expression can protect the neuron from injury induced by aluminum.

关 键 词：5-LO表达 铝 原代培养 海马神经元 腺病毒 神经元损伤 氧化应激 

分 类 号：R-332[医药卫生] R322.81