绿原酸对Aβ_(25-35)诱导的大脑皮层细胞损伤的保护作用机制  

The protective effect of chlorogenic acid on Aβ_(25-35)-induced damage to cerebral cortical neurons via macrophage activation

作　　者：周静[1] 赵宏[1] 秦书俭[1] 姚素艳[2] 郑德宇[1] 

机构地区：[1]辽宁医学院解剖学教研室 [2]辽宁医学院病理生理学教研室,辽宁锦州121001

出　　处：《中国临床解剖学杂志》2014年第3期316-320,共5页Chinese Journal of Clinical Anatomy

基　　金：国家自然科学基金(81202783);辽宁省教育厅项目(L2013335);辽宁省科技厅自然科学基金(2013022048;2013022066)

摘　　要：目的：研究绿原酸（CHA）对淀粉样β蛋白25-35（Aβ25-35）激活巨噬细胞导致大脑皮层细胞损伤的保护作用机制。方法采用生后0~3 d左右的Sprague-Dawley（SD）大鼠乳鼠，取出大脑皮层细胞然后做体外培养。培养8 d后与小鼠腹腔巨噬细胞共同培养2 d，再加入10μmol/L Aβ25-35制作阿尔茨海默病细胞模型。实验分为单独培养组、共同培养组、单独培养Aβ组、共同培养Aβ组和共同培养CHA组分别作用0.5、1、2、4、6 h后检测各组的结果。细胞损伤程度通过观察细胞形态和记数判定。p38分裂原激活蛋白激酶（p38MAPK）、有丝裂原激活蛋白激酶活化的蛋白激酶2（MAPKAPK-2）和热休克转录因子（HSP27）三种蛋白磷酸化的表达水平由Western blot检测。通过免疫荧光观察微管相关蛋白-2（MAP-2）的表达。结果 CHA显著抑制Aβ25-35处理所致的颗粒细胞密度的减少并且改善细胞的形态。Aβ25-35可以使p38MAPK的磷酸水平明显增加，在2h的时候磷酸化水平达到最高峰，4h后就逐渐下降，而使MAPKAPK-2和HSP27的磷酸化水平降低，2h降低最明显。结论 CHA通过抑制Aβ25-35诱导巨噬细胞释放炎症因子激活的p38MAPK信号转导通路，从而可以起到保护神经细胞的作用。Objective To investigate the effect of chlorogenic acid on cerebral cortical neurons injury induced by amyloid β protein via macrophage activation in vitro. Methods Cerebral cortical neurons were obtained from the cerebellar cortex of Sprague-Dawley （SD） rats and cultured. At the day 8 of culture, peritoneal macrophage of mice were added to the cerebral cortical neurons cultures and continued to be cultured for additional 2 days. Thereafter, Aβ25-35 （10 μmol/L） 10 μl were added into the co-culture of cerebral cortical neurons and macrophages. The experiment was divided into blank isolated culture group, co-culture group, isolated culture Aβ25-35 group, co-culture Aβ25-35 group and co-culture CHA-treated group for 6, 12, 24, 48, 96h, respectively. The survival of neurons and macrophages was detected by an inverted phase contrast microscope. Expressions of p38MAPK, MAPKAPK-2, HSP27 protein were tested by Western blot. Results Application of chlorogenic acid significantly improved the morphology and survival of cerebral cortical neurons damaged by Aβ25- 35-induced macrophage activation. In macrophages, the Aβ25- 35-induced p38MAPK pathway activation was inhibited by CHA at different time points. Accordingly, both the decreased expressions of phosphorylated MAPKAPK-2 and phosphorylated HSP-27 in macrophages caused by Aβ25-35 exposure were suppressed by CHA. Conclusion Chlorogenic acid is effective in reducing Aβ25-35-induced damage to cerebral cortical neurons co-cultured with macrophages. Its protective effect may be in part achieved by action against the upregulation of p38MAPK expression in macrophages.

关 键 词：绿原酸 淀粉样β蛋白25-35 大脑皮层细胞 巨噬细胞 p38分裂原激活蛋白激酶 有丝分裂原激活蛋白激酶活化的蛋白激酶2 热休克蛋白27 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]