Biochanin A attenuates LPS-induced pro-inflammatory responses and inhibits the activation of the MAPK pathway in BV2 microglial cells.
Int J Mol Med. 2015 Feb ;35(2):391-8. Epub 2014 Dec 2. PMID: 25483920
Inflammation in the brain, characterized by the activation of microglia, is believed to participate in the pathogenesis of Parkinson's disease. Biochanin A, an O-methylated isoflavone, is a natural organic compound and is classified as a phytoestrogen. In this study, using murine BV2 microglial cells, we investigated the anti-inflammatory effects of biochanin A and the possible mechanisms involved. BV2 microglial cells were treated with lipopolysaccharide (LPS) to induce pro-inflammatory responses and the cells were then treated with biochanin A. Cell viability was examined by MTT assay. The production of nitric oxide (NO) was examined using Griess reagent and intracellular reactive oxygen species (ROS production) was measured by DCFH-DA assay. The mRNA expression of interleukin-1β (IL-1β), inducible nitric oxide synthase (iNOS) and tumor necrosis factor-α (TNF-α) was examined by RT-PCR. The expression of p-ERK, p-JNK, p-p38 and iNOS was measured by western blot analysis. In addition, the protein and mRNA and phosphorylation levels of pro-inflammatory cytokines were determined by western blot analysis and RT-PCR, respectively. The results revealed that biochanin A attenuated LPS-induced microglial activation and the production of TNF‑α, IL-1β, nitric oxide and reactive oxygen species in a dose-dependent manner. Biochanin A significantly decreased the LPS-induced mRNA expression of TNF-α and IL-1β, and inhibited iNOS mRNA and protein expression. Furthermore, biochanin A significantly inhibited the LPS-induced phosphorylation of c-Jun NH2-terminal kinase (JNK), extracellular signal-regulated kinase (ERK) and p38. These findings suggest that the inhibitory effects of biochanin A on LPS-induced proinflammatory responses may be associated with the inhibition of mitogen-activated protein kinase (MAPK) signaling pathways in BV2 microglial cells.