咖啡酸对大鼠海马神经元铝盐损伤的保护作用

目的建立氯化铝致原代培养大鼠海马神经元损伤模型,观察咖啡酸对神经元铝盐损伤的保护作用。方法新生24h内SD大鼠海马神经元原代培养,7d后用免疫组化鉴定纯度;并分成空白对照组(NaCl200μmol.L-1)、铝负荷模型组(AlCl3200μmol.L-1)、铝和不同浓度的5-LO抑制剂咖啡酸(10-6、10-7、10-8mol.L-1)混合处理组。以HE染色,MTT测定,LDH漏出率,SOD活性和MDA含量为观测指标。结果神经元纯度超过95%。铝负荷致神经细胞数目明显减少,神经元突起减少变短,原代培养海马神经元活力降低,LDH漏出明显增多,SOD活性降低,MDA含量升高。咖啡酸能剂量依赖性地减轻铝盐所致的原代培养的海马神经元损伤,提高神经细胞的活力,减少LDH漏出,明显阻遏铝负荷所致的神经元SOD活性的降低和MDA含量的升高。结论咖啡酸对铝负荷所致的原代培养大鼠海马神经元损伤有一定的保护作用,其机制可能与其抑制5-LO活性,进而抑制炎症反应和氧化应激有关。

Protective effect of caffeic acid on damage induced by aluminum-overload in primary cultured rat hippocampal neuron

Aim In order to study the protective effect of caffeic acid on damage induced by aluminum-overload in primary cultured rat hippocampal neuron.Methods Primary cell cultures were obtained from the cerebral hippocampus of Newly born SD rats within 24 h.On the d 7 of neuronal culture, the immunohistory of NSE was used to identify the purity of neuron.There were 5 experimental groups,NaCl(200 μmol·L~(-1))-treated group, AlCl_3-treated group(200 μmol·L~(-1)),and aluminum+caffeic acid(10~(-6) mol·L~(-1),10~(-7) mol·L~(-1) and 10~(-8) mol·L~(-1))-treated groups. HE staining was used to observe the change of neuronal pathomorphology. The cell viability was measured by MTT assay. SOD activity, LDH leakage and MDA contents were also detected.Results The purity of neurons was more than 95%. Aluminum administration induced loss of neurons and damage to dendrite and axon.Compared with that of the control group,the decreased viability of neurons,increased leakage of LDH, decreased activity of SOD and increased contents of MDA were observed in aluminum-treated groups.Compared with that of the model group, the administration of Caffeic acid could significantly blunt the death of the primary cultured hippocampal neurons, and blunt the decrease of neuronal viability and SOD activity and the increase of LDH leakage and MDA contents.Conclusions These results suggest that caffeic acid has an obvious protective effect against neuronal damage induced by aluminum overload in primary cultured neurons. The mechanism of protection might involve the anti-inflammation and anti-oxidative effects of caffeic acid.