Inhibitory Effect and Mechanism of Fluorofenidone on the Proliferation of Rat Pulmonary Fibroblasts / 医药导报

ABSTRACT

Objective To study the inhibitory effect and mechanism of fluorofenidone on the proliferation of the rat lung fibroblasts induced by TGF-β1 . Methods Pulmonary fibroblasts cell were cultured and purified by using differentially adherent method, the third generation was used in the experiment. Cells were pretreated with 1,3,10 mmol?L-1 fluorofenidone, respectively. Pulmonary fibroblasts were identified by immunocytochemistry, and interfered by transient transfection. Cell proliferation was measured by BrdU marking. The mRNA and protein expression of eIF3α and p27 were analyzed by real-time PCR and Western blot. Results Immunocytochemistry staining identification of vimentin was positive in the third generation of pulmonary fibroblasts .TGF-β1 could obviously promote the eIF3α expression and proliferation of pulmonary fibroblasts. EIF3a siRNA was transfected with lipofectamine into cells, which obviously inhibit eIF3α expression induced by TGF-β1 , reversed cell proliferation induced by TGF-β1 , and could obviously promote p27mRNA expression induced by TGF-β1 . Fluorofenidone (3, 10 mmol?L-1 )could obviously inhibit cell proliferation induced by TGF-β1 and reduce eIF3α expression of pulmonary fibroblasts promoted by TGF-β1 . The inhibition rate was 66.7% and 78.8% respectively. Fluorofenidone (3 mmol?L-1 ,10 mmol?L-1 ) could also obviously reverse p27 expression inhibited by TGF-β1 , the reverse rate was 75. 5% and 71. 3% respectively. Conclusion eIF3α/ p27 signal pathway involves in pulmonary fibroblasts proliferation induced by TGF-β1 .Fluorofenidone can inhibit pulmonary fibroblasts proliferation induced by TGF-β1 by inhibiting eIF3α expression, enhancing p27 expression and/ or increasing p27 directly.