Anti-inflammatory effects of Viola yedoensis and the application of cell extraction methods for investigating bioactive constituents in macrophages.
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.
Korean Medicine (KM)-Application Center, Korea Institute of Oriental Medicine, 70, Cheomdanro, Dong-gu, Daegu, 41062, Republic of Korea.|College of Pharmacy, Korea University, 2511, Sejongro, Sejong, 30019, Republic of Korea.
Yun Hee
YH
Jeong
Korean Medicine (KM)-Application Center, Korea Institute of Oriental Medicine, 70, Cheomdanro, Dong-gu, Daegu, 41062, Republic of Korea.
You-Chang
YC
Oh
Korean Medicine (KM)-Application Center, Korea Institute of Oriental Medicine, 70, Cheomdanro, Dong-gu, Daegu, 41062, Republic of Korea.
Won-Kyung
WK
Cho
College of Pharmacy, Korea University, 2511, Sejongro, Sejong, 30019, Republic of Korea.
Hyeji
H
Shin
College of Pharmacy, Korea University, 2511, Sejongro, Sejong, 30019, Republic of Korea. kylee11@korea.ac.kr.
Ki Yong
KY
Lee
Korean Medicine (KM)-Application Center, Korea Institute of Oriental Medicine, 70, Cheomdanro, Dong-gu, Daegu, 41062, Republic of Korea. jyma@kiom.re.kr.
Jin Yeul
JY
Ma
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000494:pharmacology*; D002470:Cell Survival; D016207:Cytokines / Q000032:analysis / Q000378:metabolism; D051547:Heme Oxygenase-1 / Q000032:analysis / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000032:analysis / Q000378:metabolism; D016328:NF-kappa B / Q000032:analysis / Q000378:metabolism; D009569:Nitric Oxide / Q000032:analysis / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D000067996:RAW 264.7 Cells; D032413:Viola / Q000737:chemistry*
D000893:Anti-Inflammatory Agents; D016207:Cytokines; D016328:NF-kappa B; D010936:Plant Extracts; D009569:Nitric Oxide; D051547:Heme Oxygenase-1; D020928:Mitogen-Activated Protein Kinases
Bio-active components; Cell extraction; Heme oxygenase-1; Mitogen-activated protein kinase; Nuclear factor-kappaB; Viola yedoensis ethanol extract