石杉碱甲的抗炎作用及其对大鼠神经干细胞的保护作用  

Anti-inflammatory effect of huperzine A on protection of rat neural stem cells in vitro

作　　者：朱宁[1] 林继宗[2] 陈庆状 韦美丹[3] 王勇[1] 

机构地区：[1]南方医科大学珠江医院药剂科,广东广州510282 [2]中山大学第三附属医院肝胆外科,广东广州510630 [3]南方医科大学附属第三医院药剂科,广东广州510630

出　　处：《中国病理生理杂志》2013年第7期1160-1164,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30973162)

摘　　要：目的:探讨石杉碱甲(HupA)的抗炎作用及其对大鼠神经干细胞的保护作用。方法:取新生SD大鼠海马组织,分离并培养神经干细胞和小胶质细胞,建立Transwell共培养体系。将细胞分成3组:空白对照组、淀粉样β肽(Aβ)组和HupA组。Aβ组小胶质细胞层中加入终浓度为10μmol/L的Aβ1-42,HupA组于加入Aβ1-42前4 h用1μmol/L HupA预处理小胶质细胞。液相芯片技术检测炎症因子白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)和巨噬细胞炎症蛋白1α(MIP-1α)的表达,流式细胞术和Western blotting检测神经干细胞的凋亡。结果:小胶质细胞与神经干细胞共培养72 h后,与空白对照组相比,Aβ组IL-6、TNF-α和MIP-1α水平以及神经干细胞凋亡率(25.46%)均显著升高(P<0.01);HupA预处理小胶质细胞后,Aβ诱导的小胶质细胞炎症因子分泌显著减少,IL-6、TNF-α和MIP-1α的水平降低(P<0.01),同时神经干细胞凋亡率降低至8.05%(P<0.01)。Westernblotting检测结果显示,HupA组Bcl-2/Bax值显著高于Aβ组(P<0.05)。结论:石杉碱甲可以抑制小胶质细胞分泌细胞因子和趋化因子,减弱Aβ诱导的炎症反应,降低神经干细胞凋亡率,从而发挥抗炎和神经保护作用。AIM： To explore the anti-inflammatory effect of huperzine A（HupA） and its neuroprotective effect on rat neural stem cells（NSCs）.METHODS： The microglia and NSCs were isolated from neonatal rat hippocampal tissues and co-cultured in a Transwell system.The cells were divided into 3 groups： control group,amyloid beta-peptide（Aβ） group and HupA group.The microglia layer in Aβ group was treated with Aβ1-42（10 μmol/L）,while that in HupA group was pretreated with HupA（1 μmol / L） before Aβ1-42 stimulation.The culture supernatant levels of inflammatory mediators,including interleukin 6（IL-6）,tumor necrosis factor α（TNF-α） and macrophage inflammatory protein 1α（MIP1α）,were detected by LiquiChip technique.The apoptosis of NSCs was determined by flow cytometry and Western blotting.RESULTS： The microglia secreted a large number of inflammatory mediators with the stimulation of Aβ.In Aβ group,the levels of IL-6,TNF-α and MIP-1α were significantly higher than those in control group at 72 h（P 0.01）,and the apoptotic rate of NSCs was 25.46%（P 0.01）.In HupA group,the concentrations of IL-6,TNF-α and MIP-1α decreased significantly as compared with Aβ group（P 0.01）,and the apoptotic rate of NSCs was only 8.05%（P 0.01）.The Bcl-2 / Bax ratio in HupA group was higher than that in Aβ group（P 0.05）.CONCLUSION： Huperzine A reduces the secretion of cytokines and chemokines,and attenuates microglia-mediated neuroinflammation,thus protecting NSCs against inflammation-induced apoptosis.

关 键 词：石杉碱甲 小胶质细胞 神经干细胞 炎症 神经保护作用 淀粉样β肽类 

分 类 号：R363[医药卫生—病理学]