Pirfenidone inhibits proliferation, arrests the cell cycle, and downregulates heat shock protein-47 and collagen type I in rat hepatic stellate cells in vitro

Mol Med Rep. 2015 Jul;12(1):309-14. doi: 10.3892/mmr.2015.3403. Epub 2015 Mar 3.

Abstract

Pirfenidone (esbiret) is an established anti-fibrotic and anti-inflammatory drug used to treat idiopathic pulmonary fibrosis. In the present study, the dose-dependent effects of pirfenidone on the cell cycle, proliferation and expression of heat shock protein (HSP)-47 and collagen type I in a cultured rat hepatic stellate cell line (HSC-T6) were investigated. Following pirfenidone treatment, cell proliferation was determined using the cell counting kit-8 assay and the cell cycle was measured using flow cytometry. HSP-47 expression was estimated using western blot analysis and collagen type I mRNA was assessed using reverse transcription quantitative polymerase chain reaction. Pirfenidone induced significant dose-dependent inhibition of proliferation in HSC-T6 cells. Cell viability was unaffected by treatment with pirfenidone (0, 10 or 100 µM) for 24 and 72 h. However, after 24 h, HSC-T6 cells exhibited dose-dependent decreases in HSP-47 protein and collagen I mRNA levels. In conclusion, pirfenidone inhibited HSC-T6 cell proliferation, arrested the cell cycle and reduced the expression of HSP-47 and collagen type I, indicating that pirfenidone may be a promising drug in the treatment of liver fibrosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anti-Inflammatory Agents, Non-Steroidal / pharmacology*
  • Blotting, Western
  • Cell Line
  • Cell Proliferation / drug effects*
  • Cell Survival / drug effects
  • Collagen Type I / genetics*
  • Collagen Type I / metabolism
  • Down-Regulation / drug effects*
  • G1 Phase Cell Cycle Checkpoints / drug effects
  • HSP47 Heat-Shock Proteins / genetics*
  • HSP47 Heat-Shock Proteins / metabolism
  • Hepatic Stellate Cells / cytology
  • Hepatic Stellate Cells / drug effects
  • Hepatic Stellate Cells / metabolism
  • Pyridones / pharmacology*
  • RNA, Messenger / metabolism
  • Rats

Substances

  • Anti-Inflammatory Agents, Non-Steroidal
  • Collagen Type I
  • HSP47 Heat-Shock Proteins
  • Pyridones
  • RNA, Messenger
  • pirfenidone