PMID- 25890150 OWN - NLM STAT- MEDLINE DCOM- 20160309 LR - 20181113 IS - 1742-2094 (Electronic) IS - 1742-2094 (Linking) VI - 12 DP - 2015 Feb 20 TI - Transcription factor myocyte enhancer factor 2D regulates interleukin-10 production in microglia to protect neuronal cells from inflammation-induced death. PG - 33 LID - 10.1186/s12974-015-0258-z [doi] LID - 33 AB - BACKGROUND: Neuroinflammatory responses have been recognized as an important aspect in the pathogenesis of Parkinson's disease (PD). Transcriptional regulation plays a critical role in the process of inflammation. Transcription factor myocyte enhancer factor 2D (MEF2D) is identified as a central factor in transmission of extracellular signals and activation of the genetic programs in response to a wide range of stimuli in several cell types, including neurons. But its presence and function in microglia have not been reported. We therefore investigated the effect of MEF2D in activated microglia on the progress of neuroinflammation and the survival of neurons. METHODS: BV2 cells and primary cultured glial cells were stimulated with lipopolysaccharide (LPS). Samples from cells were examined for MEF2D expression, interleukin-10 (IL-10), and tumor necrosis factor alpha (TNF-alpha) by immunoblotting, quantitative real-time PCR (qPCR) or enzyme-linked immunosorbent assay (ELISA). The activity of MEF2D was examined by electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation assay (ChIP). Recombinant lentivirus expressing shRNA specific to MEF2D was used to silence MEF2D expression in BV2 cells. The role of IL-10 transcriptionally induced by MEF2D on neuronal survival was assessed by anti-IL-10 neutralizing antibody. The survival of neurons was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Male C57bl/6 mice were used to establish an acute PD model. Brain sections and cell slides were tested by immunofluorescence. RESULTS: We demonstrated that MEF2D was present in microglia. Activation of microglia was associated with an increase in MEF2D level and activity in response to different stimuli in vivo and in vitro. MEF2D bound to a MEF2 consensus site in the promoter region of IL-10 gene and stimulated IL-10 transcription. Silencing MEF2D decreased the level of IL-10, increased the TNF-alpha mRNA, and promoted inflammation-induced cytotoxicity, consistent with the result of inhibiting IL-10 activity with an anti-IL-10 neutralizing antibody. CONCLUSIONS: Our study identifies MEF2D as a critical regulator of IL-10 gene expression that negatively controls microglia inflammation response and prevents inflammation-mediated cytotoxicity. FAU - Yang, Shaosong AU - Yang S AD - Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, 710038, China. yangshaosong@aliyun.com. FAU - Gao, Li AU - Gao L AD - Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, 710038, China. gaoli089@yeah.net. FAU - Lu, Fangfang AU - Lu F AD - Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, 710038, China. misslff@163.com. FAU - Wang, Bao AU - Wang B AD - Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, 710038, China. bob.kane@163.com. FAU - Gao, Fei AU - Gao F AD - Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, 710038, China. 18092662863@163.com. FAU - Zhu, Gang AU - Zhu G AD - Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, 710038, China. zhugang.0308@163.com. FAU - Cai, Zhibiao AU - Cai Z AD - Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, 710038, China. zhibiaocai@163.com. FAU - Lai, Juan AU - Lai J AD - Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, 710038, China. laijuan_1988@163.com. FAU - Yang, Qian AU - Yang Q AD - Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, 710038, China. qianyang@fmmu.edu.cn. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150220 PL - England TA - J Neuroinflammation JT - Journal of neuroinflammation JID - 101222974 RN - 0 (Aif1 protein, mouse) RN - 0 (Calcium-Binding Proteins) RN - 0 (Lipopolysaccharides) RN - 0 (MEF2 Transcription Factors) RN - 0 (Microfilament Proteins) RN - 0 (RNA, Messenger) RN - 130068-27-8 (Interleukin-10) RN - 9P21XSP91P (1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine) RN - EC 1.14.16.2 (Tyrosine 3-Monooxygenase) SB - IM MH - 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine/pharmacology MH - Animals MH - Animals, Newborn MH - Apoptosis/drug effects/*physiology MH - Brain/cytology MH - Calcium-Binding Proteins/metabolism MH - Cells, Cultured MH - Chromatin Immunoprecipitation MH - Electrophoretic Mobility Shift Assay MH - Interleukin-10/genetics/*metabolism MH - Lipopolysaccharides/toxicity MH - MEF2 Transcription Factors/genetics/*metabolism MH - Male MH - Mice MH - Mice, Inbred C57BL MH - Microfilament Proteins/metabolism MH - Microglia/drug effects/*metabolism MH - RNA, Messenger/metabolism MH - Time Factors MH - Tyrosine 3-Monooxygenase/metabolism PMC - PMC4339472 EDAT- 2015/04/19 06:00 MHDA- 2016/03/10 06:00 PMCR- 2015/02/20 CRDT- 2015/04/19 06:00 PHST- 2014/10/09 00:00 [received] PHST- 2015/01/30 00:00 [accepted] PHST- 2015/04/19 06:00 [entrez] PHST- 2015/04/19 06:00 [pubmed] PHST- 2016/03/10 06:00 [medline] PHST- 2015/02/20 00:00 [pmc-release] AID - 10.1186/s12974-015-0258-z [pii] AID - 258 [pii] AID - 10.1186/s12974-015-0258-z [doi] PST - epublish SO - J Neuroinflammation. 2015 Feb 20;12:33. doi: 10.1186/s12974-015-0258-z.