Procyanidin B2 inhibits the activation of hepatic stellate cells and angiogenesis via the Hedgehog pathway during liver fibrosis

J Cell Mol Med. 2019 Sep;23(9):6479-6493. doi: 10.1111/jcmm.14543. Epub 2019 Jul 21.

Abstract

Background: Liver fibrosis is a wound-healing process of liver featured by the over-deposition of extracellular matrix (ECM) and angiogenesis. However, the effective treatment is lacking. Procyanidin B2 (PB2) is a flavonoid extract abundant in grape seeds with anti-oxidant, anti-inflammatory and anti-cancer properties. The present study aimed to determine effects of PB2 on liver fibrosis.

Method: The CCl4-induced mouse liver fibrosis model and a human hepatic stellate cell (HSC) line (LX2 cells) were used to study the activation, ECM production and angiogenesis of HSCs through Western blotting analysis, immunohistochemistry, immunofluorescence staining, flow cytometry and tubulogenesis assay. A Hedgehog (Hh) pathway inhibitor (cyclopamine) and Smoothened agonist (SAG) were used to investigate the role of PB2 on Hh pathway.

Results: The results showed that PB2 could inhibit the proliferation and induce apoptosis of HSCs. PB2 could also down-regulate the expressions of VEGF-A, HIF-1α, α-SMA, Col-1 and TGF-β1 of HSCs in vivo and in vitro. The application of SAG and cyclopamine proved that PB2 targets on Hh pathway.

Conclusions: PB2 inhibited the Hh pathway to suppress the activation, ECM production and angiogenesis of HSCs, therefore reverses the progression of liver fibrosis in vivo and in vitro.

Keywords: Hedgehog pathway; angiogenesis; hepatic stellate cells; liver fibrosis; procyanidin B2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biflavonoids / pharmacology*
  • Carbon Tetrachloride / pharmacology
  • Catechin / pharmacology*
  • Cell Line
  • Cell Proliferation / drug effects
  • Down-Regulation / drug effects
  • Hedgehog Proteins / metabolism*
  • Hepatic Stellate Cells / drug effects*
  • Hepatic Stellate Cells / metabolism
  • Humans
  • Liver / drug effects
  • Liver / metabolism
  • Liver Cirrhosis / drug therapy*
  • Liver Cirrhosis / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Morphogenesis / drug effects
  • Neovascularization, Pathologic / drug therapy*
  • Neovascularization, Pathologic / metabolism
  • Proanthocyanidins / pharmacology*
  • Signal Transduction / drug effects*
  • Transforming Growth Factor beta1 / metabolism
  • Veratrum Alkaloids / pharmacology

Substances

  • Biflavonoids
  • Hedgehog Proteins
  • Proanthocyanidins
  • Transforming Growth Factor beta1
  • Veratrum Alkaloids
  • procyanidin B2
  • Catechin
  • Carbon Tetrachloride
  • cyclopamine