PMID- 35172258 OWN - NLM STAT- Publisher LR - 20231019 IS - 1618-095X (Electronic) IS - 0944-7113 (Linking) VI - 99 DP - 2022 May TI - The sesquiterpene lactone estafiatin exerts anti-inflammatory effects on macrophages and protects mice from sepsis induced by LPS and cecal ligation puncture. PG - 153934 LID - S0944-7113(22)00012-5 [pii] LID - 10.1016/j.phymed.2022.153934 [doi] AB - BACKGROUND: Previously, we found that the water extract of Artermisia scoparia Waldst. & Kit suppressed the cytokine production of lipopolysaccharide (LPS)-stimulated macrophages and alleviated carrageenan-induced acute inflammation in mice. Artemisia contains various sesquiterpene lactones and most of them exert immunomodulatory activity. PURPOSE: In the present study, we investigated the immunomodulatory effect of estafiatin (EST), a sesquiterpene lactone derived from A. scoparia, on LPS-induced inflammation in macrophages and mouse sepsis model. STUDY DESIGN AND METHODS: Murine bone marrow-derived macrophages (BMDMs) and THP-1 cells, a human monocytic leukemia cell line, were pretreated with different doses of EST for 2 h, followed by LPS treatment. The gene and protein expression of pro-inflammatory cytokines interleukin (IL)-6, tumor necrosis factor (TNF)-alpha, and inducible nitric oxide synthase (iNOS) were measured by quantitative real-time polymerase chain reaction (qPCR) and Western blot analysis. The activation of nuclear factor kappa B (NF-kappaB) and mitogen-activated protein kinases (MAPKs) was also evaluated at the level of phosphorylation. The effect of EST on inflammatory cytokine production, lung histopathology, and survival rate was assessed in an LPS-induced mice model of septic shock. The effect of EST on the production of cytokines in LPS-stimulated peritoneal macrophages was evaluated by in vitro and ex vivo experiments and protective effect of EST on cecal ligation and puncture (CLP) mice was also assessed. RESULTS: The LPS-induced expression of IL-6, TNF-alpha, and iNOS was suppressed at the mRNA and protein levels in BMDMs and THP-1 cells, respectively, by pretreatment with EST. The half-maximal inhibitory concentration (IC(50)) of EST on IL-6 and TNF-alpha production were determined as 3.2 muM and 3.1 muM in BMDMs, 3 muM and 3.4 muM in THP1 cells, respectively. In addition, pretreatment with EST significantly reduced the LPS-induced phosphorylation p65, p38, JNK, and ERK in both cell types. In the LPS-induced mice model of septic shock, serum levels of IL-6, TNF-alpha, IL-1beta, CXCL1, and CXCL2 were lower in EST-treated mice than in the control animals. Histopathology analysis revealed that EST treatment ameliorated LPS-induced lung damage. Moreover, while 1 of 7 control mice given lethal dose of LPS survived, 3 of 7 EST-treated (1.25 mg/kg) mice and 5 of 7 EST-treated (2.5 mg/kg) mice were survived. Pretreatment of EST dose-dependently suppressed the LPS-induced production of IL-6, TNF-alpha and CXCL1 in peritoneal macrophages. In CLP-induced mice sepsis model, while all 6 control mice was dead at 48 h, 1 of 6 EST-treated (1.25 mg/kg) mice and 3 of 6 EST-treated (2.5 mg/kg) mice survived for 96 h. CONCLUSION: These results demonstrated that EST exerts anti-inflammatory effects on LPS-stimulated macrophages and protects mice from sepsis. Our study suggests that EST could be developed as a new therapeutic agent for sepsis and various inflammatory diseases. CI - Copyright (c) 2022 Elsevier GmbH. All rights reserved. FAU - Ahn, Jae-Hun AU - Ahn JH AD - Laboratory Animal Medicine, College of Veterinary Medicine and BK21 FOUR Program, Chonnam National University, Gwangju 61186, Republic of Korea. Electronic address: hun2wawa@hanmail.net. FAU - Song, Eun-Jung AU - Song EJ AD - Laboratory Animal Medicine, College of Veterinary Medicine and BK21 FOUR Program, Chonnam National University, Gwangju 61186, Republic of Korea. Electronic address: ejsong14@daum.net. FAU - Jung, Do-Hyeon AU - Jung DH AD - Laboratory Animal Medicine, College of Veterinary Medicine and BK21 FOUR Program, Chonnam National University, Gwangju 61186, Republic of Korea. Electronic address: jdh6221@naver.com. FAU - Kim, Yeong-Jun AU - Kim YJ AD - Laboratory Animal Medicine, College of Veterinary Medicine and BK21 FOUR Program, Chonnam National University, Gwangju 61186, Republic of Korea. Electronic address: kimyj0587@naver.com. FAU - Seo, In-Su AU - Seo IS AD - Laboratory Animal Medicine, College of Veterinary Medicine and BK21 FOUR Program, Chonnam National University, Gwangju 61186, Republic of Korea. Electronic address: insuman1@naver.com. FAU - Park, Seong-Chan AU - Park SC AD - Laboratory Animal Medicine, College of Veterinary Medicine and BK21 FOUR Program, Chonnam National University, Gwangju 61186, Republic of Korea. FAU - Jung, You-Seok AU - Jung YS AD - Laboratory Animal Medicine, College of Veterinary Medicine and BK21 FOUR Program, Chonnam National University, Gwangju 61186, Republic of Korea. Electronic address: wjddbtjr0404@naver.com. FAU - Cho, Eun-Seo AU - Cho ES AD - Department of Food Science & Technology, Chonnam National University, Gwangju, Republic of Korea. Electronic address: dmstj33@naver.com. FAU - Mo, Sang Hyun AU - Mo SH AD - Department of Food Science & Technology, Chonnam National University, Gwangju, Republic of Korea. Electronic address: msh1568@naver.com. FAU - Hong, Jung Joo AU - Hong JJ AD - National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Chungcheongbuk 28116, Republic of Korea. Electronic address: hong75@kribb.re.kr. FAU - Cho, Jeong-Yong AU - Cho JY AD - Department of Food Science & Technology, Chonnam National University, Gwangju, Republic of Korea. Electronic address: jyongcho17@jnu.ac.kr. FAU - Park, Jong-Hwan AU - Park JH AD - Laboratory Animal Medicine, College of Veterinary Medicine and BK21 FOUR Program, Chonnam National University, Gwangju 61186, Republic of Korea. Electronic address: jonpark@jnu.ac.kr. LA - eng PT - Journal Article DEP - 20220111 PL - Germany TA - Phytomedicine JT - Phytomedicine : international journal of phytotherapy and phytopharmacology JID - 9438794 SB - IM OTO - NOTNLM OT - Estafiatin OT - Inflammation OT - Lipopolysaccharide OT - Macrophages OT - Sepsis EDAT- 2022/02/17 06:00 MHDA- 2022/02/17 06:00 CRDT- 2022/02/16 20:08 PHST- 2021/09/22 00:00 [received] PHST- 2022/01/05 00:00 [revised] PHST- 2022/01/08 00:00 [accepted] PHST- 2022/02/17 06:00 [pubmed] PHST- 2022/02/17 06:00 [medline] PHST- 2022/02/16 20:08 [entrez] AID - S0944-7113(22)00012-5 [pii] AID - 10.1016/j.phymed.2022.153934 [doi] PST - ppublish SO - Phytomedicine. 2022 May;99:153934. doi: 10.1016/j.phymed.2022.153934. Epub 2022 Jan 11.