title: Withaferin A inhibits inflammatory responses induced by Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans in macrophages.
abstract: Periodontitis is a progressive chronic inflammatory disease and a major cause of tooth loss in humans. As a withanolides, withaferin A (WA) is known to exhibit strong anti‑inflammatory activity. The present study examined whether WA inhibited inflammatory responses in macrophages in response to two representative periodontal pathogens, Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans. Murine bone marrow‑derived macrophages (BMDMs) were used in this study and cytokine production in culture supernatants was measured by enzyme‑linked immunosorbent assays. Western blot analysis was performed to determine the activation of nuclear factor‑κB and mitogen‑activated protein kinases (MAPKs) and the expression of inducible nitric oxide synthase (iNOS), toll‑like receptor (TLR) 2 and TLR4. The production of nitric oxide (NO) was determined by the Griess reaction. WA treatment was shown to decrease interleukin (IL)‑6 and tumor necrosis factor (TNF)‑α production in BMDMs in response to F. nucleatum and A. actinomycetemcomitans in a dose‑dependent manner. The phosphorylation of IκB‑α and MAPKs (p38, extracellular signal‑regulated kinases and c‑Jun N‑terminal kinases) induced by F. nucleatum and A. actinomycetemcomitans was also inhibited by WA. F. nucleatum and A. actinomycetemcomitans induced iNOS expression and NO production in BMDMs, which was inhibited by WA in a dose‑dependent manner. WA also reduced endogenous and induced expression of TLR2 and TLR4 in these cells. These results suggest that WA may be a potential therapeutic agent or preventive additive for periodontitis control.
authors.affiliation: Department of Biochemistry, College of Medicine, Konyang University, Daejeon 302‑718, Republic of Korea.
authors.forename: Eui-Jeong
authors.initials: EJ
authors.lastname: Noh
authors.affiliation: Laboratory of Animal Medicine, College of Veterinary Medicine and BK21 Project Team, Chonnam National University, Gwangju 500‑757, Republic of Korea.
authors.forename: Ming-Jung
authors.initials: MJ
authors.lastname: Kang
authors.affiliation: Department of Biochemistry, College of Medicine, Konyang University, Daejeon 302‑718, Republic of Korea.
authors.forename: Yu-Jin
authors.initials: YJ
authors.lastname: Jeong
authors.affiliation: Laboratory of Animal Medicine, College of Veterinary Medicine and BK21 Project Team, Chonnam National University, Gwangju 500‑757, Republic of Korea.
authors.forename: Jun-Young
authors.initials: JY
authors.lastname: Lee
authors.affiliation: Department of Biochemistry, College of Medicine, Konyang University, Daejeon 302‑718, Republic of Korea.
authors.forename: Jung-Hwan
authors.initials: JH
authors.lastname: Park
authors.affiliation: Department of Biochemistry, College of Medicine, Konyang University, Daejeon 302‑718, Republic of Korea.
authors.forename: Hye-Jin
authors.initials: HJ
authors.lastname: Choi
authors.affiliation: Department of Biochemistry, College of Medicine, Konyang University, Daejeon 302‑718, Republic of Korea.
authors.forename: Sang-Muk
authors.initials: SM
authors.lastname: Oh
authors.affiliation: Department of Biochemistry, College of Medicine, Konyang University, Daejeon 302‑718, Republic of Korea.
authors.forename: Kyung-Bok
authors.initials: KB
authors.lastname: Lee
authors.affiliation: Department of Biochemistry, College of Medicine, Konyang University, Daejeon 302‑718, Republic of Korea.
authors.forename: Dong-Jae
authors.initials: DJ
authors.lastname: Kim
authors.affiliation: Department of Oral Pathology, School of Dentistry, Institute of Oral Bioscience, Chonbuk National University, Jeonju, North Jeolla 561‑756, Republic of Korea.
authors.forename: Ji-Ae
authors.initials: JA
authors.lastname: Shin
authors.affiliation: Department of Oral Pathology, School of Dentistry, Institute of Oral Bioscience, Chonbuk National University, Jeonju, North Jeolla 561‑756, Republic of Korea.
authors.forename: Sung-Dae
authors.initials: SD
authors.lastname: Cho
authors.affiliation: Laboratory of Animal Medicine, College of Veterinary Medicine and BK21 Project Team, Chonnam National University, Gwangju 500‑757, Republic of Korea.
authors.forename: Jong-Hwan
authors.initials: JH
authors.lastname: Park
mesh_terms: D016976:Aggregatibacter actinomycetemcomitans; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D005674:Fusobacterium Infections / Q000188:drug therapy / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology*; D016967:Fusobacterium nucleatum; D015870:Gene Expression; D008264:Macrophages / Q000187:drug effects* / Q000382:microbiology / Q000502:physiology*; D008297:Male; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D016871:Pasteurellaceae Infections / Q000188:drug therapy / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology*; D010518:Periodontitis / Q000188:drug therapy / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D051195:Toll-Like Receptor 2 / Q000378:metabolism; D051197:Toll-Like Receptor 4; D054358:Withanolides / Q000494:pharmacology*
chemical_list: D000893:Anti-Inflammatory Agents; D016207:Cytokines; D016328:NF-kappa B; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D054358:Withanolides; D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; D020928:Mitogen-Activated Protein Kinases; C009684:withaferin A