阿魏酸钠通过抑制β淀粉样蛋白1-42所致的星形胶质细胞炎症因子释放

Sodium ferulate inhibits Aβ_(1-42)-induced hippocampal neuronal injury by suppressing the inflammatory response of astrocytes

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 investigate the inhibits effect of sodium ferulate（ SF） on astrocyte-mediated inflammatory responses during Aβ-induced hippocampal neuronal injury in vitro. Method ： The primary cultured astrocytes were exposed to Aβ1-4250 nmol / L for 24 h after with and without pretreatment with various concentrations of SF（ 50,100,200 mol / L） for 6 h,as a control,Aβ1-42 was incubated in astrocyte culture medium,in the absence of cells,for 24 h. Then,astrocytic conditioned medium（ ACM） was added respectively to cultured hippocampal neurons for 48 h,the supernatants of neurons were measured the leakage of LDH,neuronal dendrite outgrowth was observed using immunostaining for the synaptophysin,Western blot was peformed to observe the level of active caspase-3 in neurons. The supernatants of astrocytes were collected and proinflammatory cytokines IL-1β,TNF-αwere determined by sandwich ELISA,NO generation was measured by Griess method. Results：cultured hippocampal neurons treated with ACM significant increase the level of active caspase-3 and leakage of LDH,and also accompanied neuronal dendrite injury,cultured astrocytes incubated with Aβ1-42 significant increase the release of IL-1β,TNF-αand NO. Astrocytes pretreatment with SF（ 50,100,200 mol / L） decrease this proinflammatory cytokines generation and also revers the changes of neurons. Conclusion： SF-mediated neuroprotection from Aβ1-42 toxicity by suppressing the inflammatory response of astrocytes.