Anti-inflammatory effects of Viola yedoensis and the application of cell extraction methods for investigating bioactive constituents in macrophages

BMC Complement Altern Med. 2016 Jun 14;16:180. doi: 10.1186/s12906-016-1142-9.


Background: Viola yedoensis (VY, Violaceae) is a popular medicinal herb used in traditional eastern medicine for treating lots of diseases, including inflammation and its related symptoms. However, the anti-inflammatory properties of VY have not been demonstrated. In the present study, we investigated the anti-inflammatory effects of VY ethanol extract (VYE) on macrophages and attempted to identify the bioactive components of VYE.

Methods: We assessed the effects of VYE on secretion of nitric oxide (NO) and inflammatory cytokines such as tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β. In addition, we explored the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, and changes in heme oxygenase (HO)-1, nuclear factor (NF)-kB, and mitogen-activated protein kinase (MAPK) signaling pathways in RAW 264.7 macrophages stimulated by lipopolysaccharide (LPS). In addition, a rapid and useful approach to identify potential bioactive components in VYE with anti-inflammatory effects was developed using murine macrophage cell extraction coupled with high-performance liquid chromatography tandem mass spectrometry (LC-MS).

Results: We found that VYE exerted anti-inflammatory activity by inhibiting the production of key inflammation mediators and related products, as well as suppression of HO-1, NF-kB, and MAPK signaling pathway activation in RAW 264.7 cells. In addition, we identified two compounds in VYE via the cell extraction method.

Conclusions: Our results revealed that VYE exerts anti-inflammatory activities and its detailed inhibitory mechanism in macrophages. Furthermore, we identified bioactive components of VYE.

Keywords: Bio-active components; Cell extraction; Heme oxygenase-1; Mitogen-activated protein kinase; Nuclear factor-kappaB; Viola yedoensis ethanol extract.

MeSH terms

  • Animals
  • Anti-Inflammatory Agents / chemistry
  • Anti-Inflammatory Agents / pharmacology*
  • Cell Survival
  • Cytokines / analysis
  • Cytokines / metabolism
  • Heme Oxygenase-1 / analysis
  • Heme Oxygenase-1 / metabolism
  • Macrophages / drug effects*
  • Macrophages / metabolism
  • Mice
  • Mitogen-Activated Protein Kinases / analysis
  • Mitogen-Activated Protein Kinases / metabolism
  • NF-kappa B / analysis
  • NF-kappa B / metabolism
  • Nitric Oxide / analysis
  • Nitric Oxide / metabolism
  • Plant Extracts / chemistry
  • Plant Extracts / pharmacology*
  • RAW 264.7 Cells
  • Viola / chemistry*


  • Anti-Inflammatory Agents
  • Cytokines
  • NF-kappa B
  • Plant Extracts
  • Nitric Oxide
  • Heme Oxygenase-1
  • Mitogen-Activated Protein Kinases