PMID- 26467650 OWN - NLM STAT- MEDLINE DCOM- 20160621 LR - 20181113 IS - 1742-2094 (Electronic) IS - 1742-2094 (Linking) VI - 12 DP - 2015 Oct 14 TI - Requirement for endogenous heat shock factor 1 in inducible nitric oxide synthase induction in murine microglia. PG - 189 LID - 10.1186/s12974-015-0406-5 [doi] LID - 189 AB - BACKGROUND: Inducible nitric oxide synthase (iNOS) makes a great contribution to host defense and inflammation. In many settings, lipopolysaccharide (LPS) induces iNOS expression through activation of the inhibitor of kappaB-alpha (IkappaB-alpha)-nuclear factor-kappaB (NF-kappaB) cascade, whereas interferon-gamma (IFN-gamma) acts through Janus kinase (JAK)-signal transducer and activator of transcription 1 (STAT1) signals. Heat shock factor 1 (HSF1), a major regulator of heat shock protein transcription, has been shown to regulate the production of pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), but it remains obscure whether and how HSF1 affects iNOS induction. METHODS: Western blot was used to measure the protein expression. The mRNA level was measured by real-time PCR. Silence of HSF1 was achieved by small interfering RNA. Nitric oxide (NO) content and NF-kappaB binding activity were assayed by commercial kits. Chromatin immunoprecipitation (ChIP) was used to measure the binding activity of NF-kappaB and STAT1 to iNOS promoters. RESULTS: HSF1 inhibition or knockdown prevented the LPS- and/or IFN-gamma-stimulated iNOS protein expression in cultured microglia. HSF1 inhibition blocked iNOS mRNA transcription. These inhibitory effects of HSF1 inhibition on iNOS expression were confirmed in brain tissues from endotoxemic mice. Further analysis showed that HSF1 inhibition had no effect on IkappaB-alpha degradation and NF-kappaB or STAT1 phosphorylation in LPS/IFN-gamma-stimulated cells. The nuclear transport of active NF-kappaB or STAT1 was also not affected by HSF1 inhibition, but HSF1 inhibition reduced the binding of NF-kappaB and STAT1 to their DNA elements. In addition, HSF1 inhibition reduced NF-kappaB and STAT1 bindings to iNOS promoter inside the LPS/IFN-gamma-stimulated cells. CONCLUSIONS: This preventing effect of HSF1 inhibition on iNOS mRNA transcription presents the necessary role of HSF1 in iNOS induction. FAU - Huang, Chao AU - Huang C AD - Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu Province, 226001, China. 5887218@qq.com. AD - Key Laboratory of Inflammation and Molecular Drug Targets of Jiangsu Province, Nantong University, #19 Qixiu Road, Nantong, Jiangsu Province, 226001, China. 5887218@qq.com. FAU - Lu, Xu AU - Lu X AD - Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu Province, 226001, China. 418796584@qq.com. AD - Key Laboratory of Inflammation and Molecular Drug Targets of Jiangsu Province, Nantong University, #19 Qixiu Road, Nantong, Jiangsu Province, 226001, China. 418796584@qq.com. FAU - Tong, Lijuan AU - Tong L AD - Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu Province, 226001, China. 815964117@qq.com. AD - Key Laboratory of Inflammation and Molecular Drug Targets of Jiangsu Province, Nantong University, #19 Qixiu Road, Nantong, Jiangsu Province, 226001, China. 815964117@qq.com. FAU - Wang, Jili AU - Wang J AD - Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu Province, 226001, China. 372899487@qq.com. AD - Key Laboratory of Inflammation and Molecular Drug Targets of Jiangsu Province, Nantong University, #19 Qixiu Road, Nantong, Jiangsu Province, 226001, China. 372899487@qq.com. FAU - Zhang, Wei AU - Zhang W AD - Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu Province, 226001, China. asicampa@aliyun.com. AD - Key Laboratory of Inflammation and Molecular Drug Targets of Jiangsu Province, Nantong University, #19 Qixiu Road, Nantong, Jiangsu Province, 226001, China. asicampa@aliyun.com. FAU - Jiang, Bo AU - Jiang B AD - Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu Province, 226001, China. 532059984@qq.com. AD - Key Laboratory of Inflammation and Molecular Drug Targets of Jiangsu Province, Nantong University, #19 Qixiu Road, Nantong, Jiangsu Province, 226001, China. 532059984@qq.com. FAU - Yang, Rongrong AU - Yang R AD - Department of Anesthesiology, Affiliated Hospital of Nantong University, Jiangsu Province, #20Xisi Road, Nantong, Jiangsu Province, 226001, China. 172916178@qq.com. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20151014 PL - England TA - J Neuroinflammation JT - Journal of neuroinflammation JID - 101222974 RN - 0 (Aminopyridines) RN - 0 (DNA-Binding Proteins) RN - 0 (Heat Shock Transcription Factors) RN - 0 (Hsf1 protein, mouse) RN - 0 (Indazoles) RN - 0 (Lipopolysaccharides) RN - 0 (N2-(1H-indazole-5-yl)-N6-methyl-3-nitropyridine-2,6-diamine) RN - 0 (NF-kappa B) RN - 0 (RNA, Messenger) RN - 0 (RNA, Small Interfering) RN - 0 (STAT1 Transcription Factor) RN - 0 (Stat1 protein, mouse) RN - 0 (Transcription Factors) RN - 82115-62-6 (Interferon-gamma) RN - EC 1.14.13.39 (Nitric Oxide Synthase Type II) SB - IM MH - Aminopyridines/pharmacology MH - Animals MH - Animals, Newborn MH - Cell Survival/drug effects/physiology MH - Cells, Cultured MH - DNA-Binding Proteins/antagonists & inhibitors/genetics/*metabolism MH - Dose-Response Relationship, Drug MH - Heat Shock Transcription Factors MH - Indazoles/pharmacology MH - Interferon-gamma/pharmacology MH - Lipopolysaccharides/pharmacology MH - Male MH - Mice MH - Mice, Inbred C57BL MH - Microglia/drug effects/*metabolism MH - NF-kappa B/metabolism MH - Nitric Oxide Synthase Type II/genetics/*metabolism MH - Protein Binding/drug effects MH - RNA, Messenger/metabolism MH - RNA, Small Interfering/pharmacology MH - STAT1 Transcription Factor/metabolism MH - Signal Transduction/drug effects MH - Transcription Factors/antagonists & inhibitors/genetics/*metabolism PMC - PMC4607096 EDAT- 2015/10/16 06:00 MHDA- 2016/06/22 06:00 PMCR- 2015/10/14 CRDT- 2015/10/16 06:00 PHST- 2015/06/18 00:00 [received] PHST- 2015/09/28 00:00 [accepted] PHST- 2015/10/16 06:00 [entrez] PHST- 2015/10/16 06:00 [pubmed] PHST- 2016/06/22 06:00 [medline] PHST- 2015/10/14 00:00 [pmc-release] AID - 10.1186/s12974-015-0406-5 [pii] AID - 406 [pii] AID - 10.1186/s12974-015-0406-5 [doi] PST - epublish SO - J Neuroinflammation. 2015 Oct 14;12:189. doi: 10.1186/s12974-015-0406-5.