title: Effect of 15-Deoxy-△(12,14)-prostaglandin J2 on Expression of Macrophage Migration Inhibitory Factor in Mouse Monocyte/macrophage Cell Line J774A.1.
abstract: Objective To investigate the effect of 15-Deoxy-△(12,14)-prostaglandin J2 (15 d-PGJ2) on the expression of macrophage migration inhibitory factor (MIF) and its underlying mechanism in J774A.1. Methods The murine monocyte/macrophage cell line J774A.1 were divided into six groups:lipopolysaccharide (LPS) group,incubated with 1 μg/ml LPS for 1 h;normal control group,incubated with PBS for 1 h;negative control group,incubated with 5 μmol/L 15 d-PGJ2 for 1 h;15 d-PGJ2 group,incubated with 5 μmol/L 15 d-PGJ2 for 1 h followed by 1 μg/ml LPS for 1 h;GW9662 group,incubated with 5 μmol/L 15 d-PGJ2 for 1 h following GW9662 10 μmol/L for 1 h,and then incubated with 1 μg/ml LPS for 1 h;and Vehicle group,control of GW9662,GW9662 was replaced by its solvent DMSO. The expression of MIF was detected via immunofluorescence and agarose gel electrophoresis. RT-qPCR and Western blotting were used to test whether 15 d-PGJ2 could regulate mRNA and protein expression of MIF in J774A.1 upon LPS challenge. The effect of peroxisome proliferator-activated receptor-γ (PPAR-γ) antagonist GW9662 on the regulation of MIF by 15 d-PGJ2 was observed. The effects of 15 d-PGJ2 on the nuclear translocation of PPAR-γ upon LPS challenge were detected via high content screening analysis. Results MIF DNA and protein expressions were detected in J774A.1. MIF mRNA expression was up-regulated (1.75±0.09,P=0.037) when challenged with LPS and 15 d-PGJ2 inhibited its upregulation (0.84±0.08,P=0.026) in J774A.1. The protein level was consistent with the mRNA level. PPAR-γ antagonist GW9662 reversed the effect of 15 d-PGJ2 (mRNA,1.48±0.06,P=0.016;protein,1.28). Furthermore,nuclear translocation of PPAR-γ was regulated by 15 d-PGJ2 in J774A.1 upon LPS challenge(1.39±0.02 vs. 1.01±0.03,P=0.003). Conclusion 15 d-PGJ2 may down-regulate the MIF expression in J774A.1 in a PPAR-γ-dependent manner.
authors.affiliation: Municipal Laboratory for Liver Protection and Regulation of Regeneration,Department of Cell Biology,Capital Medical University,Beijing 100069,China.
authors.forename: Wei-Yang
authors.initials: WY
authors.lastname: Li
authors.affiliation: Municipal Laboratory for Liver Protection and Regulation of Regeneration,Department of Cell Biology,Capital Medical University,Beijing 100069,China.
authors.forename: Yu-Meng
authors.initials: YM
authors.lastname: Shi
authors.affiliation: Municipal Laboratory for Liver Protection and Regulation of Regeneration,Department of Cell Biology,Capital Medical University,Beijing 100069,China.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.affiliation: Municipal Laboratory for Liver Protection and Regulation of Regeneration,Department of Cell Biology,Capital Medical University,Beijing 100069,China.
authors.forename: Lin
authors.initials: L
authors.lastname: Yang
authors.affiliation: Municipal Laboratory for Liver Protection and Regulation of Regeneration,Department of Cell Biology,Capital Medical University,Beijing 100069,China.
authors.forename: Li-Ying
authors.initials: LY
authors.lastname: L I
mesh_terms: D000813:Anilides / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D019746:Intramolecular Oxidoreductases / Q000378:metabolism*; D008070:Lipopolysaccharides; D008263:Macrophage Migration-Inhibitory Factors / Q000378:metabolism*; D051379:Mice; D009000:Monocytes / Q000187:drug effects*; D047495:PPAR gamma / Q000037:antagonists & inhibitors; D015230:Prostaglandin D2 / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: C097240:15-deoxy-delta(12,14)-prostaglandin J2; C457499:2-chloro-5-nitrobenzanilide; D000813:Anilides; D008070:Lipopolysaccharides; D008263:Macrophage Migration-Inhibitory Factors; D047495:PPAR gamma; D019746:Intramolecular Oxidoreductases; C076231:Mif protein, mouse; D015230:Prostaglandin D2