Inhibitory effect of phloretin and biochanin A on IgE-mediated allergic responses in rat basophilic leukemia RBL-2H3 cells

Life Sci. 2013 Sep 17;93(9-11):401-8. doi: 10.1016/j.lfs.2013.07.019. Epub 2013 Jul 29.

Abstract

Aims: Anti-allergic effects and action mechanism of phloretin (Phl) and biochanin A (BioA) on the IgE-antigen complex-mediated allergic responses in rat basophilic leukemia RBL-2H3 cells were investigated.

Main methods: Cell viability, formation of reactive oxygen species (ROS), DPPH radical-scavenging activity, β-hexosaminidase release, production of interleukin (IL)-4, IL-13, and tumor necrosis factor-alpha (TNF-α) and phosphorylation of Akt and mitogen-activated protein kinase (MAPK) were determined by MTT assay, 2,7-dichlorofluorescein diacetate (DCF-DA) assay, DPPH radical-scavenging assay, reverse transcriptase polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA) and western blot analysis, respectively.

Key findings: Ph1 and BioA dose-dependently inhibited the formation of ROS and the release of β-hexosaminidase from the RBL-2H3 cells and also showed DPPH radical-scavenging activity. Ph1 and BioA suppressed the antigen-induced phosphorylation of the downstream signaling intermediates, including MAPK and Akt, which are critical for the production of pro-inflammatory cytokines, and also significantly attenuated the production of IgE-mediated pro-inflammatory cytokines, such as IL-4, IL-13, and TNF-α.

Significance: Phloretin and biochanin A attenuate the degranulation and allergic cytokine production through inhibition of intracellular ROS production and the phosphorylation of Akt and the MAPKs, such as ERK1/2, p38, and JNK. The results of this study suggested that these two plant flavonoids may have potent anti-allergic activity in vitro.

Keywords: Anti-allergic activity; Cytokine; Degranulation; Flavonoids; RBL-2H3 cells.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Cell Line, Tumor
  • Cytokines / drug effects
  • Cytokines / metabolism
  • Dose-Response Relationship, Drug
  • Enzyme-Linked Immunosorbent Assay
  • Free Radical Scavengers / administration & dosage
  • Free Radical Scavengers / pharmacology
  • Genistein / administration & dosage
  • Genistein / pharmacology*
  • Immunoglobulin E / immunology*
  • Leukemia, Basophilic, Acute / immunology*
  • Mitogen-Activated Protein Kinases / metabolism
  • Phloretin / administration & dosage
  • Phloretin / pharmacology*
  • Phosphorylation / drug effects
  • Proto-Oncogene Proteins c-akt / metabolism
  • Rats
  • Reactive Oxygen Species / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Cytokines
  • Free Radical Scavengers
  • Reactive Oxygen Species
  • Immunoglobulin E
  • Genistein
  • Proto-Oncogene Proteins c-akt
  • Mitogen-Activated Protein Kinases
  • Phloretin
  • biochanin A