Fluorofenidone inhibits apoptosis of renal tubular epithelial cells in rats with renal interstitial fibrosis

This study aimed to investigate the mechanism of fluorofenidone (AKF-PD) in treating renal interstitial fibrosis in rats with unilateral urinary obstruction (UUO). Thirty-two male Sprague-Dawley rats were randomly divided into sham, UUO, UUO + enalapril, and UUO + AKF-PD groups. All rats, except sham, underwent left urethral obstruction surgery to establish the animal model. Rats were sacrificed 14 days after surgery, and serum was collected for renal function examination. Kidneys were collected to observe pathological changes. Immunohistochemistry was performed to assess collagen I (Col I) protein expression, and terminal deoxynucleotidyl transferase-mediated nick end-labeling staining to observe the apoptosis of renal tubular epithelial cells. The expression of Fas-associated death domain (FADD), apoptotic protease activating factor-1 (Apaf-1), and C/EBP homologous protein (CHOP) proteins was evaluated by immunohistochemistry and western blot analysis. AKF-PD showed no significant effect on renal function in UUO rats. The pathological changes were alleviated significantly after enalapril or AKF-PD treatment, but with no significant differences between the two groups. Col I protein was overexpressed in the UUO group, which was inhibited by both enalapril and AKF-PD. The number of apoptotic renal tubular epithelial cells was much higher in the UUO group, and AKF-PD significantly inhibited epithelial cells apoptosis. The expression of FADD, Apaf-1, and CHOP proteins was significantly upregulated in the UUO group and downregulated by enalapril and AKF-PD. In conclusion, AKF-PD improved renal interstitial fibrosis by inhibiting apoptosis of renal tubular epithelial cells in rats with UUO.

Effect of fluorofenidone on renal interstitial fibrosis in rats with unilateral ureteral obstruction / 中南大学学报(医学版)

Objective:To investigate the effect of fluorofenidone on renal interstitial fibrosis in rats with unilateral ureteral obstruction (UUO) and to observe the effect of fluorofenidone on the expressions of collagen type Ⅰ (Col Ⅰ),collagen type Ⅲ (Col Ⅲ),α-smooth muscle actin (α-SMA),connective tissue growth factor (CTGF),platelet derived growth factor (PDGF) in the renal tissues of UUO rats.Methods:Male Sprague-Dawley (SD) rats were randomly divided into a sham-operated group,a UUO group,and a flurofenidone group (n=5).UUO model was induced by ligating the left ureter in rats.The rats were treated with 125 mg/(kg.d) fluorofenidone by gastric gavage in the fluorofenidone group at 24 h before the operation,and the rats were treated with the identical dose of 0.5％ sodium carboxyl methyl cellulose (CMC-Na) in the other 2 groups.The rats were sacrificed at 14 days after UUO.Pathological changes of the renal tissue were observed by HE and Masson staining,the mRNA expressions of Col Ⅰ,Col Ⅲ,α-SMA,PDGF and CTGF were detected by real-time PCR,and the protein expressions of Col Ⅰ,Col Ⅲ,PDGF and CTGF were detected by immunohistochemical staining.Results:The renal interstitial damage index,relative collagen area and mRNA and protein expressions of Col Ⅰ and Col Ⅲ in the renal tissues of the rats in the UUO group significantly increased (P＜0.05),and fluorofenidone could reduce these indexes (P＜0.05).Compared with the sham-operated group,the protein expressions ofα-SMA,PDGF,CTGF and the mRNA expressions of PDGF and CTGF in the renal tissues of the rats in the UUO group were increased,but fluorofenidone could decrease the protein expressions of α-SMA,PDGF,CTGF and the mRNA expressions of PDGF and CTGF (P＜0.05).Conclusion:Fluorofenidone (125 mg/kg·d) could attenuate renal interstitial fibrosis through inhibition offibroblast proliferation,myofibroblastic activation,PDGF and CTGF expression.

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

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.