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目的:研究阿魏酸钠对Aβ1-42激活的培养星形胶质细胞引起的海马神经细胞损伤的保护作用。方法:原代培养海马神经细胞与星形胶质细胞,星形胶质细胞用阿魏酸钠(50,100,200 mol/L)预处理6 h后,加入50 nmol/L的Aβ1-42作用24 h,将作用后的星形胶质细胞条件培养液(astrocytic conditioned medium,ACM)加入到培养海马神经细胞中作用48 h,以加入Aβ1-42的无细胞培养液为对照。ELISA法测定星形胶质细胞培养液中IL-1β、TNF-α表达,Griess法测定培养液中NO的含量;测定各组海马神经元乳酸脱氢酶(LDH)释放、采用突触素荧光免疫组织化学染色观察神经元损伤、Western蛋白印迹法检测神经元caspase-3蛋白表达。结果:与正常对照组相比,星形胶质细胞培养液加入Aβ1-42处理后IL-1β、TNF-α、NO生成量明显增加,海马神经元细胞加入ACM后突触素减少,LDH漏出量增加,磷酸化的caspase-3蛋白表达明显增加;应用阿魏酸钠(50,100,200 mol/L)预处理6 h可明显对抗Aβ1-42引起的星形胶质细胞及海马神经元细胞的这些改变。结论:阿魏酸钠通过抑制星形胶质细胞炎症因子释放对抗Aβ1-42引起的海马神经元损伤。
AIM: To study the protective effect of sodium ferulate on hippocampal neuronal injury induced by Aβ 1-42 -activated astrocytes. Methods: Primary cultured hippocampal neurons and astrocytes were pretreated with sodium ferulate (50, 100, 200 mol / L) for 6 h, then treated with 50 nmol / L Aβ1-42 for 24 h, The treated astrocytic conditioned medium (ACM) was added to the cultured hippocampal neurons for 48 h, and the cell-free culture medium containing Aβ1-42 was used as a control. ELISA was used to determine the expression of IL-1β and TNF-α in astrocyte culture medium. The content of NO in culture fluid was determined by Griess method. The release of lactate dehydrogenase (LDH) Neuronal damage was observed by immunohistochemical staining. Caspase-3 protein expression in neurons was detected by Western blotting. Results: Compared with the normal control group, the production of IL-1β, TNF-α and NO significantly increased after Aβ1-42 treatment, and decreased in the hippocampal neurons after addition of ACM The phosphorylation of caspase-3 protein was significantly increased. Pretreatment with sodium ferulate (50, 100, 200 mol / L) for 6 h significantly inhibited these changes of astrocytes and hippocampal neurons induced by Aβ1-42 . CONCLUSION: Sodium ferulate antagonizes hippocampal neuronal injury induced by Aβ1-42 by inhibiting the release of astrocyte inflammatory cytokines.