摘要
目的观察知母皂苷(SAaB)是否能对抗淀粉样β蛋白片段25-35(Aβ25-35)引起的巨噬细胞炎症介质释放并探讨其信号转导通路的影响。方法小鼠腹腔巨噬细胞培养24 h,加入不同浓度SAaB(10、30和100μmol·L-1)或加入不同的阻断剂(诱导性一氧化氮合酶阻断剂SMT、MEK1的特异性阻断剂PD98059、p38MAPK特异性阻断剂SB203580和PI3K特异性阻断剂LY294002),之后加入Aβ25-35(20μmol·L-1)继续培养。Aβ25-35作用2 h后,采用Westernblot观察巨噬细胞磷酸化Akt/PKB蛋白表达水平的改变。Aβ25-35作用48 h后,取巨噬细胞培养上清液分别测定肿瘤坏死因子-α(TNF-α)和一氧化氮(NO)的含量变化。结果 PD98059和LY294002均可明显抑制Aβ25-35引起的巨噬细胞TNF-α产生增加和磷酸化Akt/PKB表达明显增加。SMT可完全对抗Aβ25-35引起的NO释放增加。SAaB可明显抑制Aβ25-35引起的TNF-a和NO的产生增加,并呈明显的浓度依赖性。SAaB也可明显抑制Aβ25-35引起磷酸化Akt/PKB表达增加。结论 SAaB能够明显的抑制Aβ25-35引起的巨噬细胞炎症因子释放,这种抑制作用部分通过SAaB抑制Akt/PKB信号转导通路产生。
Aim To investigate the inhibitory effect and the possible signaling mechanism of Sapoin from Anemarrhena asphdeloids Bge（SAaB） on the release of inflammatory mediators induced by amyloid β-protein fragments 25-35（Aβ25-35） in cultured macrophages.Methods Cultured mouse peritoneal macrophages were preincubated with SAaB（10,30 and 100 μmol·L^-1） or specific kinase inhibitors including SMT,PD98059,SB20358 and LY294002（20 μmol·L-1 Aβ25-35）,then stimulated with Aβ25-35（20 μmol·L^-1）.After stimulation with Aβ25-35 for the indicated time,total cellular extracts were prepared for Western blot analysis of Akt/PKB.The supernatants of macrophages were collected and analyzed for tumor necrosis factor-α（TNF-α） and nitric oxide（NO） generation.Results Aβ25-35 significantly induced increase in phospho-Akt/PKB protein expression without affecting total protein levels and the production of TNF-a and NO in cultured macrophages.iNOS inhibitor SMT notablely reduced Aβ25-35-induced increase of NO generation.PD98059 and LY294002 could distinctly inhibit the production of TNF-α and the expression of phospho-Akt/PKB.SAaB（100 μmol·L^-1） significantly suppressed Aβ25-35-induced increase in phospho-Akt/PKB protein level.In addition,SAaB（10,30 and 100 μmol·L^-1） also decreased TNF-α and NO generation in supernatants of cultured macrophages in a concentration-dependent manner.Conclusion SAaB can significantly inhibit the over-release of inflammatory mediators induced by Aβ25-35 in cultured macrophages,a process when the down regulation of Akt/PKB signal transduction pathway took part in.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2011年第5期695-700,共6页
Chinese Pharmacological Bulletin
基金
辽宁省教育厅创新团队项目(NoLT2010064)
辽宁医学院院内资助课题(NoY2010Z003)
作者简介
刘卓（1979-），女，硕士．讲师．研究方向：抗老年痴呆药物，E—mail：lyliuzhuo200732@yahoo．com．cn；
金英（1960-），女，博士．教授，研究方向：神经药理学及信号传导，通讯作者．Tel／1：Fax：0416-4673409，E—mail：jyjinying@yahoo．com.cn
