川芎嗪通过抑制RAGE-ERK1/2-p38-NFκB信号通路及活性氧生成改善阿尔茨海默病大鼠脑组织炎性  

Tetramethylpyrazine Protect Rats against the Inflammation and Aβ25-35 induced ROS by Targeting the RAGE-ERK1/2-p38-NFκB Pathway

作　　者：刘长安[1] 朱洁[1] 蔡标[1] 黄金玲[1,2] 

机构地区：[1]安徽中医药大学中西医结合临床学院,合肥230038 [2]安徽省中医药科学院中西医结合研究所,合肥230038

出　　处：《中国药学杂志》2014年第13期1126-1132,共7页Chinese Pharmaceutical Journal

基　　金：安徽省高等学校省级优秀青年人才基金项目(2012SQRL103);安徽中医药大学自然科学研究基金项目(2013zr007);安徽省高校省级自然科学研究项目(KJ2013Z160)

摘　　要：目的观察川芎嗪对阿尔茨海默病大鼠脑组织炎性及原代海马神经元的RAGE-ERK1/2-p38-NFκB信号转导系统的影响。方法采用Aβ25-35双侧海马注射建立阿尔茨海默病大鼠模型,观察川芎嗪对阿尔茨海默病大鼠海马组织RAGE蛋白表达及脑组织炎症因子水平;原代培养大鼠海马神经元细胞,并转染大鼠RAGE基因过表达载体,加入Aβ25-35建立体外细胞模型,四甲基偶氮唑蓝法检测细胞活性,DCFH-DA荧光染料检测活性氧,Real-time qPCR检测RAGE mRNA表达,Western blot检测ERK1/2、p38、IκB磷酸化水平。结果川芎嗪可显著性降低阿尔茨海默病大鼠海马组织RAGE蛋白表达,显著降低脑组织炎症因子白细胞介素-1β、白细胞介素-6、肿瘤坏死因子-α水平;川芎嗪可显著提高原代海马神经元细胞存活,显著抑制活性氧生成,降低RAGE mRNA水平,显著降低ERK1/2、p38、IκB的磷酸化水平。当海马神经元过表达RAGE后,川芎嗪的相关作用可以被逆转。结论川芎嗪通过对阿尔茨海默病大鼠海马神经元RAGE-ERK1/2-p38-NF-κB信号通路的下调作用,有效抑制神经元活性氧生成,降低Aβ的神经毒性,改善阿尔茨海默病大鼠脑组织炎性,进而发挥其抗阿尔茨海默病及神经保护作用。OBJECITVE To investigate the effects of tetramethylpyrazine on inflammatory response and RAGE mediated signal transduction in hippocampus of rats with Alzheimer＇s disease induced by Aβ25-35and explored potential mechanism. METHODS Rats received a hibateral intracerebroventricular fusion of Aβ25-35. Tetramethylpyrazine was administrated through i. v. injection at 20 and 60 mg · kg-1 · d-1 for 21 d. The expression of RAGE and inflammatory factors（ including IL-1β, IL-6,TNF-α）were assessed immunohis- toehemieal and ELISA, respectively. Further, an AD cell model （ primary cultured hippocampal neurons of SD neonatal rats ） and a RAGE-over expressing cell model（primary cultured hippocampal neurons transfected with pCMV/hygro-RAGE） were used for investi- gating the mechanisms of tetramethylpyrazine＇s effects. Cell viability was determined by MTT assay and intracellular reactive oxygen species （ROS） was identified by DCFH-DA staining. Real time fluorescent quantitative PCR and Western blot method were employed to detect the transcriptional level（RAGE） and phosphorylation of targeted proteins （including ERK1/2, p38, hoB ） in all groups. RE- SULTS Tetramethylpyrazine decreased RAGE expression（ P 〈 0. 05 ） in hippoeampus and attenuated the inflammatory factors inclu- ding IL-1β （ P 〈 0. 05 ）, IL-6 （ P 〈 0. 05 ） , and TNF-α （ P 〈 0. 05 ） in brain tissue of Aβ25-35-treated rats. Incubation with tetrameth- ylpyrazine attenuated the decrease of survival rate of primary cultured hippoeampal neurons （compared with model group, P 〈 0. 05 ）and reduced Aβ25-35 induced ROS （ compared with model group, P 〈 0. 05 ）. CONCLUSION Meanwhile, tetramethylpyrazine significant- ly inhibited the upregulation of RAGE transcription and markedly depressed the activation of extracellular sigual-regulated kinase 1/2 （ERK1/2） -p38 mitogen-activated protein kinase （p38MAPK） -nuclear factor KB （NFKB） pathway and the downstream NFKB inflam- matory response subsequent t

关 键 词：川芎嗪 阿尔茨海默病 活性氧 RAGE—ERKl 2-p38-NF-kB信号通路 炎症反应 

分 类 号：R965[医药卫生—药理学]