PMID- 28662496 OWN - NLM STAT- MEDLINE DCOM- 20171107 LR - 20190212 IS - 1421-9778 (Electronic) IS - 1015-8987 (Linking) VI - 42 IP - 3 DP - 2017 TI - Macrophage Inflammatory Protein-2 in High Mobility Group Box 1 Secretion of Macrophage Cells Exposed to Lipopolysaccharide. PG - 913-928 LID - 10.1159/000478646 [doi] AB - BACKGROUND/AIMS: Macrophage inflammatory protein-2 (MIP-2), a type of leukocyte chemokine, is primarily produced by macrophages, and levels increase significantly in early inflammation. However, the precise biological functions and mechanisms of MIP-2 in the development of inflammation remain unclear. The purposes of the present study were to investigate the role of MIP-2 in inflammation induced by lipopolysaccharide (LPS) in vitro and to determine the possibility of blocking the high mobility group box 1 (HMGB1) signalling pathway via MIP-2 inhibition. METHODS: Macrophage cells (RAW264.7, U937 and THP-1 cells) were divided into control and treatments groups. Expression levels of interleukin-6 (IL-6), interleukin-1beta (IL-1beta), tumour necrosis factor-alpha (TNF-alpha), HMGB1, chemokine (C-C motif) ligand-2 (Ccl-2), Toll-like receptor-4 (TLR-4), inducible nitric oxide synthase (iNOS), phosphorylated MAPKs (p38, ERKs, JNKs), PI3K/Akts, JAKs/STAT3, IkappaB, and cytoplasmic and nuclear NF-kappaB p65 in RAW264.7 cells were detected by qRT-PCR, enzyme-linked immunosorbent assay (ELISA) or western blot assays. RESULTS: mip-2 siRNA and an anti-MIP-2 antibody significantly reduced the expression levels of Ccl-2, TLR-4, iNOS, IL-6, IL-1beta, HMGB1, and TNF-alpha in RAW264.7 cells exposed to LPS (P<0.01). Additionally, mRNA expression levels of HMGB1 and TLR-4 in cells treated with LPS+mip-2 siRNA were significantly lower than those in cells treated with LPS alone (P<0.01 or P<0.05). The MIP-2 antibody significantly suppressed activation of p38-MAPK, p-STAT3, and p-Akts and translocation of NF-kappaB p65 from the cytoplasm to the nucleus in RAW264.7 exposed to LPS (P<0.01 or P<0.05). CONCLUSION: mip-2 siRNA and the MIP-2 antibody can reduce the inflammatory effects induced by LPS in macrophage cells. The mechanisms may occur through down-regulation of p38-MAPK, STAT3 and Akts phosphorylation and translocation of NF-kappaB p65. MIP-2 plays an important role in inflammation induced by LPS. CI - (c) 2017 The Author(s). Published by S. Karger AG, Basel. FAU - Chaochao, Qin AU - Chaochao Q AD - State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou, China. FAU - Lou, Guohua AU - Lou G AD - State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou, China. FAU - Yang, Ying AU - Yang Y AD - State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou, China. FAU - Liu, Yanning AU - Liu Y AD - State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou, China. FAU - Hu, Ying AU - Hu Y AD - State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou, China. FAU - Min, Zheng AU - Min Z AD - State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou, China. FAU - Chen, Ping AU - Chen P AD - State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou, China. FAU - He, Jiliang AU - He J AD - Department of Environmental Medicine, School of Public Health, Zhejiang University, Hangzhou, China. FAU - Chen, Zhi AU - Chen Z AD - State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou, China. LA - eng PT - Journal Article DEP - 20170626 PL - Germany TA - Cell Physiol Biochem JT - Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology JID - 9113221 RN - 0 (Chemokine CXCL2) RN - 0 (HMGB1 Protein) RN - 0 (HMGB1 protein, mouse) RN - 0 (Interleukin-1beta) RN - 0 (Interleukin-6) RN - 0 (Lipopolysaccharides) RN - 0 (RNA, Small Interfering) RN - 0 (Toll-Like Receptor 4) RN - 0 (Tumor Necrosis Factor-alpha) RN - EC 1.14.13.39 (Nitric Oxide Synthase Type II) SB - IM MH - Animals MH - Chemokine CXCL2/genetics/*immunology MH - Down-Regulation MH - Gene Expression Regulation MH - HMGB1 Protein/genetics/*immunology MH - Inflammation/genetics/*immunology MH - Interleukin-1beta/genetics MH - Interleukin-6/genetics MH - Lipopolysaccharides/*immunology MH - Macrophages/*immunology/metabolism MH - Mice MH - Nitric Oxide Synthase Type II/genetics MH - RAW 264.7 Cells MH - *RNA Interference MH - RNA, Small Interfering/genetics MH - Toll-Like Receptor 4/genetics MH - Tumor Necrosis Factor-alpha/genetics OTO - NOTNLM OT - Acute inflammation OT - HMGB1 OT - Macrophage inflammatory protein-2 OT - Macrophages EDAT- 2017/07/01 06:00 MHDA- 2017/11/08 06:00 CRDT- 2017/06/30 06:00 PHST- 2016/09/06 00:00 [received] PHST- 2017/04/07 00:00 [accepted] PHST- 2017/07/01 06:00 [pubmed] PHST- 2017/11/08 06:00 [medline] PHST- 2017/06/30 06:00 [entrez] AID - 000478646 [pii] AID - 10.1159/000478646 [doi] PST - ppublish SO - Cell Physiol Biochem. 2017;42(3):913-928. doi: 10.1159/000478646. Epub 2017 Jun 26.