title: Proteomic Analysis of HDAC3 Selective Inhibitor in the Regulation of Inflammatory Response of Primary Microglia.
abstract: HDAC3 has been shown to regulate inflammation. However, the role of HDAC3 in primary microglia is largely unknown. RGFP966 is a newly discovered selective HDAC3 inhibitor. In this study, we used protein mass spectrometry to analyze protein alterations in LPS-treated primary microglia with the application of RGFP966. Generally, about 2000 proteins were studied. 168 of 444 (37.8%) LPS-induced proteins were significantly reduced with the treatment of RGFP966, which mainly concentrated on Toll-like receptor signaling pathway. In this regard, we selected Toll-like receptor 2 (TLR2), TLR3, TLR6, MAPK p38, CD36, and spleen tyrosine kinase (SYK) for further validation and found that they were all significantly upregulated after LPS stimulation and downregulated in the presence of RGFP966. Additionally, RGFP966 inhibited supernatant tumor necrosis factor (TNF)-αand Interleukin 6 (IL-6) concentrations. Activation of STAT3 and STAT5 was partially blocked by RGFP966 at 2 h after LPS-stimulation. The fluorescence intensity of CD16/32 was significantly decreased in LPS + RGFP966-treated group. In conclusion, our data provided a hint that RGFP966 may be a potential therapeutic medication combating microglia activation and inflammatory response in central nervous system, which was probably related to its repressive impacts on TLR signaling pathways and STAT3/STAT5 pathways.
authors.affiliation: Department of Neurology, Affiliated Drum Tower Hospital, Nanjing University Medical school, Nanjing, Jiangsu 210008, China.
authors.forename: Mingxu
authors.identifier: 0000-0001-8460-4482
authors.initials: M
authors.lastname: Xia
authors.affiliation: Department of Neurology, Affiliated Drum Tower Hospital, Nanjing University Medical school, Nanjing, Jiangsu 210008, China.
authors.forename: Qiuchen
authors.initials: Q
authors.lastname: Zhao
authors.affiliation: Department of Neurology, Affiliated Drum Tower Hospital, Nanjing University Medical school, Nanjing, Jiangsu 210008, China.
authors.forename: He
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Neurology, Affiliated Drum Tower Hospital, Nanjing University Medical school, Nanjing, Jiangsu 210008, China.
authors.forename: Yanting
authors.initials: Y
authors.lastname: Chen
authors.affiliation: The State Key Laboratory of Brain and Cognitive Sciences, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Zengqiang
authors.initials: Z
authors.lastname: Yuan
authors.affiliation: Department of Neurology, Affiliated Drum Tower Hospital, Nanjing University Medical school, Nanjing, Jiangsu 210008, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Neurology, Affiliated Drum Tower Hospital, Nanjing University Medical school, Nanjing, Jiangsu 210008, China.
authors.forename: Meijuan
authors.identifier: 0000-0002-5375-5349
authors.initials: M
authors.lastname: Zhang
mesh_terms: D000178:Acrylamides / Q000494:pharmacology* / Q000627:therapeutic use; D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism; D004660:Encephalitis / Q000139:chemically induced / Q000188:drug therapy / Q000378:metabolism*; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D006655:Histone Deacetylases / Q000378:metabolism*; D008070:Lipopolysaccharides; D051379:Mice; D008810:Mice, Inbred C57BL; D017628:Microglia / Q000187:drug effects* / Q000378:metabolism*; D010655:Phenylenediamines / Q000494:pharmacology* / Q000627:therapeutic use; D040901:Proteomics; D050796:STAT3 Transcription Factor / Q000378:metabolism; D050799:STAT5 Transcription Factor; D051195:Toll-Like Receptor 2 / Q000378:metabolism; D000099186:Histone Deacetylase 3
chemical_list: D000178:Acrylamides; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases; D008070:Lipopolysaccharides; D010655:Phenylenediamines; D050796:STAT3 Transcription Factor; D050799:STAT5 Transcription Factor; D051195:Toll-Like Receptor 2; D000099186:Histone Deacetylase 3; C000603861:RGFP966; C494087:Stat3 protein, mouse