绿原酸对Aβ25-35诱导的小脑颗粒细胞损伤的保护作用机制

神经药理学报 ›› 2013, Vol. 3 ›› Issue (6): 9-14.

绿原酸对Aβ_25-35诱导的小脑颗粒细胞损伤的保护作用机制

周静¹ ，赵宏¹ ，刘建生¹ ，姚素艳² ，郑德宇¹

1 江西科技师范大学生命科学学院,南昌,330013,中国
2 江西中医药大学附属医院,南昌,330006,中国

出版日期:2013-12-26 发布日期:2014-06-27
通讯作者: 郑德宇，男，博士，硕士生导师；研究方向：神经退行性疾病和骨组织工程修复骨缺损；E-mail: 857367505@qq.com
作者简介:周静，女，研究生；研究方向：神经生物学；E-mail:371494095@qq.com
基金资助:
辽宁省教育厅项目(No.L2013335)，辽宁省科技厅自然科学基金(No.2013022048、No.2013022066)

Influence of Chlorogenic Acid on Aβ_25-35-induced Damage to Granular Cell in Cerebellum

ZHOU Jing¹, ZHAO Hong¹, LIU Jian-sheng¹, YAO Su-yan², ZHENG De-yu¹

1 Department of Anatomy, Liaoning Medical College, Jinzhou, 121001, China
2 Department of Pathophysiology, Liaoning Medical College, Jinzhou, 121001, China

Online:2013-12-26 Published:2014-06-27
Contact: 郑德宇，男，博士，硕士生导师；研究方向：神经退行性疾病和骨组织工程修复骨缺损；E-mail: 857367505@qq.com
About author:周静，女，研究生；研究方向：神经生物学；E-mail:371494095@qq.com
Supported by:
辽宁省教育厅项目(No.L2013335)，辽宁省科技厅自然科学基金(No.2013022048、No.2013022066)

摘要/Abstract

摘要： 目的：探讨绿原酸(chlorogenic acid，CHA)对淀粉样β蛋白25-35（amyloid β-protein25-35，Aβ_25-35）激活小鼠巨噬细胞引起小脑颗粒细胞（cerebellar granular cell，CGC）损伤的保护作用机制。方法：实验分为空白对照组、Aβ_25-35处理组和Aβ_25-35+CHA共同处理不同时间组；空白对照组：加入等量培养基；Aβ_25-35损伤组：加入Aβ_25-3510 μmol·L^-1；Aβ_25-35和 CHA处理不同时间组：将10 μmol·L^-1 Aβ_25-35和1000 μmol·L^-1 CHA加入共同培养2天的小脑颗粒细胞和巨噬细胞，分别作用6、12、24、48、96 h。应用倒置相差显微镜观察小脑颗粒细胞数和形态变化并计数；应用Western blot检测p38分裂原激活蛋白激酶(p38 mitogen-activated protein kinase，p38MAPK)、下游底物有丝裂原激活蛋白激酶活化的蛋白激酶2（mitogen-activated protein kinase activating protein kinase-2, MAPKAPK-2)、热休克转录因子（heat-shock protein27, HSP27）三种蛋白的磷酸化表达的改变。结果：CHA保护组的小脑颗粒细胞数量较Aβ_25-35损伤组明显增加；CHA作用96 h可明显抑制Aβ_25-35引起的巨噬细胞磷酸化的p38MAPK表达明显增加，逆转Aβ_25-35引起的巨噬细胞磷酸化的MAPKAPK-2和磷酸化的HSP27表达降低的变化。结论：绿原酸是通过抑制Aβ_25-35激活巨噬细胞p38MAPK信号通路的蛋白表达起到保护小脑颗粒细胞的作用。

关键词: 绿原酸, 淀粉样&beta, 蛋白, 小脑颗粒细胞, 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

Abstract: Objective: To investigate the effects of chlorogenic acid on granular cell in cerebellum induced by amyloid β- protein -induced macrophage activation in vitro. Methods: The experiment was divided into blank control group, Aβ_25-35 treatment group and Aβ_25-35+CHA co-processing of different time group; Blank control group: Equal amount of culture medium; Aβ_25-35 treatment group: with Aβ_25-3510 μmol·L^-1; Aβ_25-35and different time of CHA treatment group; The co-cultured system of cerebellar granular cells and macrophages were cultivated for 2 days and then the 10 μmol·L^-1 Aβ_25-35 and 1000 μmol·L^-1 CHA were added into this system for 6, 12, 24, 48, 96 h respectively. The growth phynotype of neurons and macrophages were detected by inverted phase contrast microscope; The expressions of p38MAPK, MAPKAPK-2, HSP27 protein were tested by Western blot. Results: The numbers cerebellar granule cell of chlorogenic acid protection group was increased significantly compared with Aβ_25-35 injury group. CHA could inhibite increased expression of phosphorylated p38MAPK of macrophage induced by Aβ_25-35 at difference time; Decreased expression of phosphorylated MAPKAPK-2 and phosphorylated HSP-27of macrophage induced by Aβ_25-35 could increased after the 1000 μmol·L^-1 CHA added into the medium. Conclusion: The protection of Chlorogenic acid for cerebellar granular cells induced by Aβ_25-35 via inhibition the activation of p38MAPK signaling pathway, which decreased the expression of p38MAPK and increased the expression of MAPKAPK-2, HSP-27.

Key words: chlorogenic acid, A&beta, _25-35, CGC, macrophage, p38MAPK, MAPKAPK-2, HSP27

周静, 赵宏, 刘建生, 姚素艳, 郑德宇. 绿原酸对Aβ_25-35诱导的小脑颗粒细胞损伤的保护作用机制[J]. 神经药理学报, 2013, 3(6): 9-14.

ZHOU Jing, ZHAO Hong, LIU Jian-sheng, YAO Su-yan, ZHENG De-yu. Influence of Chlorogenic Acid on Aβ_25-35-induced Damage to Granular Cell in Cerebellum[J]. ACTA NEUROPHARMACOLOGICA, 2013, 3(6): 9-14.

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