title: Methylene blue increases the amount of HSF1 through promotion of PKA-mediated increase in HSF1-p300 interaction.
abstract: Heat shock factor 1 (HSF1) critically contributes to the host defense, and its amount determines the mobilization efficiency of HSF1 under stress conditions. To date, how HSF1 amount is regulated remains largely unknown. We found that methylene blue (MB), an anti-oxidative and anti-inflammatory agent, increased the amount of HSF1 in BV-2 microglia, primary microglia, astrocytes, neurons and vital organs. The increased HSF1 contributed to a more increase in nuclear translocation of HSF1, association of HSF1 with heat shock protein 70 (Hsp70) promoters and Hsp70 expression levels, and also induced a synergistic protection against oxidative stress-induced injuries in MB and heat shock-treated cells. The MB-induced increase in the amount of HSF1 was not associated with light exposition as well as the change in HSF1 gene transcription or macroautophagy, but associated with the proteasome-ubiquitin system. The acetyltransferase p300 was considered to mediate the effect of MB on HSF1, as p300 inhibition or silencing prevented the increase in HSF1-p300 interaction as well as the amount and acetylation level of HSF1 in MB-treated cells. Moreover, inhibition of protein kinase A α (PKAα) was found to attenuate the MB-induced increase in HSF1 amount and HSF1-p300 interaction. These findings were ascertained in primary microglia, astrocytes and neurons where p300 or PKA inhibition prevented the increase in the amount of HSF1 after MB treatment. Taken together, our results showed that MB increases the amount of HSF1 through promotion of PKA-mediated increase in HSF1-p300 interaction, providing evidence to illustrate a new pharmacological effect of MB in clinical application.
authors.affiliation: Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu, 226001, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Huang
authors.affiliation: Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu, 226001, China.
authors.forename: Wenfeng
authors.initials: W
authors.lastname: Hu
authors.affiliation: Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu, 226001, China.
authors.forename: Jili
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu, 226001, China.
authors.forename: Lijuan
authors.initials: L
authors.lastname: Tong
authors.affiliation: Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu, 226001, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Lu
authors.affiliation: Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu, 226001, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Wu
authors.affiliation: Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu, 226001, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Ling
authors.affiliation: Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu, 226001, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Jiang
authors.affiliation: Department of Pharmacology, School of Pharmacy, Nantong University, #19 Qixiu Road, Nantong, Jiangsu, 226001, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Department of Anesthesiology, Affiliated Hospital of Nantong University, Nantong, Jiangsu, 226001, China.
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Pain Management, Wuhan Pu-Ai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430033, China.
authors.forename: Qiuju
authors.initials: Q
authors.lastname: Xiong
authors.affiliation: Invasive Technology Department, Nantong First People's Hospital, the Second Affiliated Hospital of Nantong University, #6 North Road Hai'er Xiang, Nantong, Jiangsu, 226001, China.
authors.forename: Yibin
authors.initials: Y
authors.lastname: Qin
authors.affiliation: Invasive Technology Department, Nantong First People's Hospital, the Second Affiliated Hospital of Nantong University, #6 North Road Hai'er Xiang, Nantong, Jiangsu, 226001, China. Electronic address: charliechao888888@163.com.
authors.forename: Rongrong
authors.initials: R
authors.lastname: Yang
mesh_terms: D000107:Acetylation; D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D050881:E1A-Associated p300 Protein / Q000378:metabolism*; D000076249:Heat Shock Transcription Factors; D008751:Methylene Blue / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D017628:Microglia / Q000187:drug effects* / Q000378:metabolism*; D008954:Models, Biological; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D009928:Organ Specificity; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D059748:Proteolysis / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D004268:DNA-Binding Proteins; D000076249:Heat Shock Transcription Factors; C490017:Hsf1 protein, mouse; D047428:Protein Kinase Inhibitors; D014157:Transcription Factors; D050881:E1A-Associated p300 Protein; C494361:Ep300 protein, mouse; D017868:Cyclic AMP-Dependent Protein Kinases; D008751:Methylene Blue
keywords: HSF1; Hsp70; MB; PKAα; p300