X Chen1, Y Shen1, H Zhao1, W Guo2. 1. Myopic Laser Treatment Center, Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010059, China. 2. Department of Emergency Medicine, Second Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010091, China.
Abstract
OBJECTIVE: To investigate the molecular mechanism by which pirfenidone inhibits scar formation through the TGF-β/Smad pathway. METHODS: Cultured rabbit tenon fibroblasts (RTFs) were treated with different concentrations of pirfenidone to determine its initial active concentration and optimum concentration of pirfenidone for inhibiting RTF proliferation using CCK-8 assay. In RTFs treated with pirfenidone at the initial and optimal concentrations, expressions of TGF-β3, collagen I and collagen III were examined with both immunofluorescence assay and Western blotting, and their mRNA expression levels were detected using RT-PCR. RESULTS: The initial and optimal concentrations of pirfenidone for inhibiting RTF proliferation were 0.1 mg/mL and 0.27 mg/mL, respectively. In RTFs treated with pirfenidone at the two concentrations for 24 h, both immunofluorescence assay and Western blotting showed significantly lowered protein expressions of TGF-β3, collagen I and collagen III as compared with those in the control group (P < 0.05). The mRNA expressions of TGF-β3, collagen I and collagen III in the RTFs were also significantly lowered after treatment with pirfenidone at the initial and optimal concentrations (P < 0.05). CONCLUSIONS: Pirfenidone concentration-dependently inhibits the proliferation of RTFs possibly by down-regulating the expression of TGF-β3 in the TGF-β/Smad pathway.
OBJECTIVE: To investigate the molecular mechanism by which pirfenidone inhibits scar formation through the TGF-β/Smad pathway. METHODS: Cultured rabbit tenon fibroblasts (RTFs) were treated with different concentrations of pirfenidone to determine its initial active concentration and optimum concentration of pirfenidone for inhibiting RTF proliferation using CCK-8 assay. In RTFs treated with pirfenidone at the initial and optimal concentrations, expressions of TGF-β3, collagen I and collagen III were examined with both immunofluorescence assay and Western blotting, and their mRNA expression levels were detected using RT-PCR. RESULTS: The initial and optimal concentrations of pirfenidone for inhibiting RTF proliferation were 0.1 mg/mL and 0.27 mg/mL, respectively. In RTFs treated with pirfenidone at the two concentrations for 24 h, both immunofluorescence assay and Western blotting showed significantly lowered protein expressions of TGF-β3, collagen I and collagen III as compared with those in the control group (P < 0.05). The mRNA expressions of TGF-β3, collagen I and collagen III in the RTFs were also significantly lowered after treatment with pirfenidone at the initial and optimal concentrations (P < 0.05). CONCLUSIONS: Pirfenidone concentration-dependently inhibits the proliferation of RTFs possibly by down-regulating the expression of TGF-β3 in the TGF-β/Smad pathway.
Authors: David Alejandro Lopez-de la Mora; Cibeles Sanchez-Roque; Margarita Montoya-Buelna; Sergio Sanchez-Enriquez; Silvia Lucano-Landeros; Jose Macias-Barragan; Juan Armendariz-Borunda Journal: Int J Med Sci Date: 2015-10-14 Impact factor: 3.738