Effect of 15-Deoxy-△(12,14)-prostaglandin J2 on Expression of Macrophage Migration Inhibitory Factor in Mouse Monocyte/macrophage Cell Line J774A.1.
10.3881/j.issn.1000-503X.2016.03.001
- Author:
Wei-Yang LI
1
;
Yu-Meng SHI
1
;
Xin LIU
1
;
Lin YANG
1
;
Li-Ying L I
1
Author Information
1. Municipal Laboratory for Liver Protection and Regulation of Regeneration,Department of Cell Biology,Capital Medical University,Beijing 100069,China.
- Publication Type:Journal Article
- MeSH:
Anilides;
pharmacology;
Animals;
Cell Line;
Intramolecular Oxidoreductases;
metabolism;
Lipopolysaccharides;
Macrophage Migration-Inhibitory Factors;
metabolism;
Mice;
Monocytes;
drug effects;
PPAR gamma;
antagonists & inhibitors;
Prostaglandin D2;
analogs & derivatives;
pharmacology
- From:
Acta Academiae Medicinae Sinicae
2016;38(3):247-252
- CountryChina
- Language:English
-
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.
