title: The suppressive effects of gx-50 on Aβ-induced chemotactic migration of microglia.
abstract: Microglia, the main immune cells of the central nervous system (CNS), play a vital role in the development of AD. Once microglia are activated, they migrate to neuritic plaques and persistently release pro-inflammatory mediators that lead to neuroinflammation and neuronal degeneration, accelerating the progression of AD. In this study, we analyzed whether an AD candidate drug, N-[2-(3,4-dimethoxyphenyl)ethyl]-3-phenyl-acrylamide (gx-50), a compound extracted from Sichuan pepper (Zanthoxylum bungeanum), exhibited suppressive effects on the chemotactic migration of microglia induced by Aβ. At first, the effects of gx-50 on the migration of primary cultured microglia to Aβ were detected by transwell assay, and the secretion of chemokine CCL5 was measured by ELISA assay. Then, the release of TGF-β1 was detected by ELISA and quantitative real-time PCR, and the activation of the TGF-β1-Smad2 pathway was analyzed by Western blotting. The LDH assay revealed that cell viability was not affected by gx-50 at concentrations from 0.01 to 100 μM; thus, combined with our previous studies, 1 μM was chosen as the treatment concentration. The cell transwell measurement demonstrated that gx-50 suppressed the chemotactic migration of microglia by nearly 50% and inhibited the increase in CCL5 triggered by Aβ. Moreover, the analysis of the TGF-β1-Smad2 pathway revealed that gx-50 can antagonize Aβ-induced down-regulation of TGF-β1 at both the mRNA and protein levels and stimulate the signal pathway activation. Simultaneously, gx-50 pretreatment also significantly enhanced the phosphorylation of glycogen synthase kinase-3β (GSK-3β), which correlated closely with the migration of microglia. In conclusion, in the presence of Aβ, gx-50 pretreatment inhibited the excessive chemotactic migration of microglia.
authors: School of Life Science and Biotechnology, Shanghai Jiao Tong University, No. 800, Dongchuan Road, Minhang District, Shanghai 200240, China.
authors: Yubing
authors: Y
authors: Guo
authors: School of Life Science and Biotechnology, Shanghai Jiao Tong University, No. 800, Dongchuan Road, Minhang District, Shanghai 200240, China.
authors: Shi
authors: S
authors: Shi
authors: School of Life Science and Biotechnology, Shanghai Jiao Tong University, No. 800, Dongchuan Road, Minhang District, Shanghai 200240, China.
authors: Maoping
authors: M
authors: Tang
authors: School of Life Science and Biotechnology, Shanghai Jiao Tong University, No. 800, Dongchuan Road, Minhang District, Shanghai 200240, China.
authors: Dongli
authors: D
authors: Liang
authors: School of Life Science and Biotechnology, Shanghai Jiao Tong University, No. 800, Dongchuan Road, Minhang District, Shanghai 200240, China.
authors: Wangjie
authors: W
authors: Xu
authors: School of Life Science and Biotechnology, Shanghai Jiao Tong University, No. 800, Dongchuan Road, Minhang District, Shanghai 200240, China.
authors: Lianyun
authors: L
authors: Wang
authors: School of Life Science and Biotechnology, Shanghai Jiao Tong University, No. 800, Dongchuan Road, Minhang District, Shanghai 200240, China. Electronic address: zhaoxiaw@sjtu.edu.cn.
authors: Zhaoxia
authors: Z
authors: Wang
authors: School of Life Science and Biotechnology, Shanghai Jiao Tong University, No. 800, Dongchuan Road, Minhang District, Shanghai 200240, China. Electronic address: zdqiao@sjtu.edu.cn.
authors: Zhongdong
authors: Z
authors: Qiao
mesh_terms: D000178:Acrylamides / Q000494:pharmacology*; D016229:Amyloid beta-Peptides; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002633:Chemotaxis / Q000187:drug effects*; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D017628:Microglia / Q000187:drug effects* / Q000502:physiology; D010446:Peptide Fragments; D051381:Rats; D051899:Smad2 Protein / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism*
chemical_list: D000178:Acrylamides; D016229:Amyloid beta-Peptides; D010446:Peptide Fragments; D051899:Smad2 Protein; C497784:Smad2 protein, rat; D053773:Transforming Growth Factor beta1; C075222:amyloid beta-protein (1-42); C583359:lemairamin; D000071679:Glycogen Synthase Kinase 3 beta; C000605508:Gsk3b protein, rat; D038362:Glycogen Synthase Kinase 3
keywords: Alzheimer's disease; Chemotaxis; Microglia; TGF-β1-Smad2 pathway