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Macrophage migration inhibitory factor enhances lipopolysaccharide-induced fibroblast proliferation by inducing
Zheng-de Xi1, Chang-Yi Xie2, Ye-Bin Xi2
1Shanghai Institute of Immunology, Department of Immunology and Microbiology, Shanghai Jiao Tong University School of Medicine, Room 905, Building 5, 280 South Shanghai Chongqing Road, Shanghai, 200025, China. xizhengde@sjtu.edu.cn.
Background:
Fibroblast proliferation is a common manifestation of chronic inflammatory diseases, including rheumatoid arthritis (RA), Crohn's disease and ulcerative colitis, etc. To alleviate patient suffering, the mechanism underlying fibroblast proliferation should be elucidated.
Methods:
CCK-8 assay was used to assess the stimulatory effect of LPS and macrophage migration inhibitory factor (MIF) on fibroblast proliferation. Then, TLR4 expression on fibroblast cell membrane was carried out by confocal scanning microscopy. Finally, real-time fluorescent quantitative PCR and flow cytometry were applied to determine the expression of TLR4 after MIF challenge.
Results:
LPS alone directly stimulated the fibroblast proliferation. In addition, MIF showed co-stimulatory effect on LPS-induced fibroblast proliferation. Interestingly, fibroblast overtly expressed TLR4 without stimulation. After MIF stimulation, real-time PCR showed TLR4 mRNA levels were increased by about 33% in the fibroblasts; in agreement, TLR4 expression on the fibroblast membrane was increased by about 20%, as shown by flow cytometry.
Conclusions:
These findings indicated MIF elevates TLR4 expression in fibroblast, enhancing LPS-induced cell proliferation.
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