title: Ethyl acetate extract from Asparagus cochinchinensis exerts anti‑inflammatory effects in LPS‑stimulated RAW264.7 macrophage cells by regulating COX‑2/iNOS, inflammatory cytokine expression, MAP kinase pathways, the cell cycle and anti-oxidant activity.
abstract: Asparagus cochinchinesis (A. cochinchinesis) is a medicine traditionally used to treat fever, cough, kidney disease, breast cancer, inflammatory disease and brain disease in northeast Asian countries. Although numerous studies of the anti‑inflammatory effects of A. cochinchinesis have been conducted, the underlying mechanisms of such effects in macrophages remain to be demonstrated. To investigate the mechanism of suppressive effects on the inflammatory response in macrophages, alterations of the nitric oxide (NO) level, the cell viability, inducible nitric oxide synthase (iNOS) and cyclooxygenase‑2 (COX‑2) expression levels, inflammatory cytokine expression, the mitogen-activated protein kinase (MAPK) signaling pathway, cell cycle arrest and reactive oxygen species (ROS) levels were measured in lipopolysaccharide (LPS)-activated RAW264.7 cells following treatment with ethyl acetate extract from A. cochinchinesis root (EaEAC). RAW264.7 cells pretreated two different concentrations of EaEAC prior to LPS treatment exhibited no significant toxicity. The concentration of NO was significantly decreased in the EaEAC + LPS treated group compared with the vehicle + LPS treated group. A similar decrease in mRNA transcript level of COX‑2, iNOS, pro-inflammatory cytokines [tumor necrosis factor‑α and interleukin (IL)‑1β] and anti‑inflammatory cytokines (IL‑6 and IL‑10) was detected in the EaEAC + LPS treated group compared with the vehicle + LPS treated group, although the decrease rate varied. Enhancement of the phosphorylation of MAPK family members following LPS treatment was partially rescued in the EaEAC pretreated group, and the cell cycle was arrested at the G2/M phase. Furthermore, the EaEAC pretreated group exhibited a reduced level of ROS generation compared with the vehicle + LPS treated group. Taken together, these results suggest that EaEAC suppresses inflammatory responses through inhibition of NO production, COX‑2 expression and ROS production, as well as differential regulation of inflammatory cytokines and cell cycle in RAW264.7 cells. In addition, these results provide strong evidence to suggest that EaEAC may be considered as an important candidate for the treatment of particular inflammatory diseases.
authors.affiliation: College of Natural Resources and Life Science/Life and Industry Convergence Research Institute, Pusan National University, Miryang, Gyeongsangnamdo 627‑706, Republic of Korea.
authors.forename: Hyun Ah
authors.initials: HA
authors.lastname: Lee
authors.affiliation: College of Natural Resources and Life Science/Life and Industry Convergence Research Institute, Pusan National University, Miryang, Gyeongsangnamdo 627‑706, Republic of Korea.
authors.forename: Eun Kyoung
authors.initials: EK
authors.lastname: Koh
authors.affiliation: College of Natural Resources and Life Science/Life and Industry Convergence Research Institute, Pusan National University, Miryang, Gyeongsangnamdo 627‑706, Republic of Korea.
authors.forename: Ji Eun
authors.initials: JE
authors.lastname: Sung
authors.affiliation: College of Natural Resources and Life Science/Life and Industry Convergence Research Institute, Pusan National University, Miryang, Gyeongsangnamdo 627‑706, Republic of Korea.
authors.forename: Ji Eun
authors.initials: JE
authors.lastname: Kim
authors.affiliation: College of Natural Resources and Life Science/Life and Industry Convergence Research Institute, Pusan National University, Miryang, Gyeongsangnamdo 627‑706, Republic of Korea.
authors.forename: Sung Hwa
authors.initials: SH
authors.lastname: Song
authors.affiliation: College of Natural Resources and Life Science/Life and Industry Convergence Research Institute, Pusan National University, Miryang, Gyeongsangnamdo 627‑706, Republic of Korea.
authors.forename: Dong Seob
authors.initials: DS
authors.lastname: Kim
authors.affiliation: College of Natural Resources and Life Science/Life and Industry Convergence Research Institute, Pusan National University, Miryang, Gyeongsangnamdo 627‑706, Republic of Korea.
authors.forename: Hong Joo
authors.initials: HJ
authors.lastname: Son
authors.affiliation: Gangrim Organics, Miryang, Gyeongsangnamdo 627‑706, Republic of Korea.
authors.forename: Chung Yeoul
authors.initials: CY
authors.lastname: Lee
authors.affiliation: College of Human Ecology, Pusan National University, Busan 609‑735, Republic of Korea.
authors.forename: Hee Seob
authors.initials: HS
authors.lastname: Lee
authors.affiliation: Biologics Division, Ministry of Food and Drug Safety, Cheongju, Chungcheongbukdo 361‑951, Republic of Korea.
authors.forename: Chang Joon
authors.initials: CJ
authors.lastname: Bae
authors.affiliation: College of Natural Resources and Life Science/Life and Industry Convergence Research Institute, Pusan National University, Miryang, Gyeongsangnamdo 627‑706, Republic of Korea.
authors.forename: Dae Youn
authors.initials: DY
authors.lastname: Hwang
mesh_terms: D000085:Acetates / Q000737:chemistry; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D000975:Antioxidants / Q000378:metabolism; D027761:Asparagus Plant / Q000737:chemistry*; D002453:Cell Cycle / Q000187:drug effects*; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism*; D016166:Free Radical Scavengers / Q000494:pharmacology; D018836:Inflammation Mediators / Q000378:metabolism; D008070:Lipopolysaccharides; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D009569:Nitric Oxide / Q000096:biosynthesis; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism*; D010936:Plant Extracts / Q000494:pharmacology*; D000067996:RAW 264.7 Cells; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D000085:Acetates; D000893:Anti-Inflammatory Agents; D000975:Antioxidants; D016207:Cytokines; D016166:Free Radical Scavengers; D018836:Inflammation Mediators; D008070:Lipopolysaccharides; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D009569:Nitric Oxide; C007650:ethyl acetate; D052247:Nitric Oxide Synthase Type II; D051546:Cyclooxygenase 2