title: Antifibrotic effects of pirfenidone on Tenon's fibroblasts in glaucomatous eyes: comparison with mitomycin C and 5-fluorouracil.
abstract: OBJECTIVE
  The purpose of this study was to evaluate the antifibrotic effects of pirfenidone (PFD) on primary cultured human Tenon's fibroblasts (HTFs) from primary open-angle glaucoma (POAG) eyes, compared to mitomicin C (MMC) and 5-fluorouracil (5-FU).
  
  
  METHODS
  Samples of human Tenon's capsule were obtained during respective surgeries from three groups of patients: patients with cataract (CAT group), patients with POAG who underwent glaucoma filtration surgery (GFS) (POAG1 group), and patients with POAG who underwent GFS due to failed bleb of previous GFS (POAG2 group). Cell toxicity, cell migration, and the expression level of α-smooth muscle actin (α-SMA) protein were evaluated in primary cultured HTFs from the three patient groups after treatment (PFD, MMC, or 5-FU).
  
  
  RESULTS
  Overall, cell viability after PFD treatment was higher compared to MMC treatment (82.3 ± 5.1 % vs 56.7 ± 3.8 %; p = 0.001) and comparable to 5-FU treatment (82.3 ± 5.1 % vs 85.7 ± 10.7 %, p = 0.214) at the same concentration (0.4 mg/ml). Both 0.3 mg/ml PFD and 0.1 mg/ml MMC inhibited cell migration compared to control (without treatment) cells (p = 0.014 and 0.005, respectively), while 0.2 mg/ml 5-FU showed the highest degree of cell migration among the three agents in the POAG1 group (PFD vs MMC vs 5-FU; 29.5 ± 2.1 % vs 34.5 ± 0.7 % vs 76.0 ± 8.5 %, PFD vs MMC; p = 1.000, PFD vs 5-FU; p = 0.008, MMC vs 5-FU; p = 0.011). PFD (0.1 or 0.3 mg/ml) and MMC (0.05 and 0.1 mg/ml) treatment significantly reduced the protein expression level of α-SMA in the POAG 1 group (all p < 0.05), and the α-SMA protein level following treatment with 0.3 mg/ml PFD was lower than that of 0.1 mg/ml MMC (p = 0.040).
  
  
  CONCLUSIONS
  PFD showed less cytotoxicity compared to MMC. PFD and MMC inhibited cell migration and reduced α-SMA protein expression levels, while 5-FU showed neither inhibition of cell migration nor reduction in α-SMA expression level. These findings indicate PFD as a potential adjunctive antifibrotic agent to prevent bleb failure during GFS.
authors.affiliation: St. Mary's Eye Center, Seoul, Korea.
authors.forename: Jung Hwa
authors.initials: JH
authors.lastname: Na
authors.forename: Kyung Rim
authors.initials: KR
authors.lastname: Sung
authors.forename: Jin A
authors.initials: JA
authors.lastname: Shin
authors.forename: Jung Il
authors.initials: JI
authors.lastname: Moon
mesh_terms: D000199:Actins / Q000378:metabolism; D000328:Adult; D000477:Alkylating Agents / Q000494:pharmacology; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D015153:Blotting, Western; D002386:Cataract / Q000473:pathology; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D005472:Fluorouracil / Q000494:pharmacology*; D005902:Glaucoma, Open-Angle / Q000473:pathology* / Q000601:surgery; D006801:Humans; D008297:Male; D016685:Mitomycin / Q000494:pharmacology*; D011728:Pyridones / Q000494:pharmacology*; D058475:Tenon Capsule / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D014130:Trabeculectomy; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; D000477:Alkylating Agents; D000894:Anti-Inflammatory Agents, Non-Steroidal; D011728:Pyridones; D014409:Tumor Necrosis Factor-alpha; D016685:Mitomycin; C093844:pirfenidone; D005472:Fluorouracil