PMID- 25283329 OWN - NLM STAT- MEDLINE DCOM- 20150806 LR - 20181202 IS - 1873-4847 (Electronic) IS - 0955-2863 (Linking) VI - 25 IP - 12 DP - 2014 Dec TI - Involvement of de novo ceramide synthesis in pro-inflammatory adipokine secretion and adipocyte-macrophage interaction. PG - 1309-16 LID - S0955-2863(14)00172-7 [pii] LID - 10.1016/j.jnutbio.2014.07.008 [doi] AB - Interaction between adipocytes and macrophages has been suggested to play a central role in the pathogenesis of obesity. Ceramide, a sphingolipid de novo synthesized from palmitate, is known to stimulate pro-inflammatory cytokine secretion from multiple types of cells. To clarify whether de novo synthesized ceramide contributes to cytokine dysregulation in adipocytes and macrophages, we observed cytokine secretion in mature 3T3-L1 adipocytes (L1) and RAW264.7 macrophages (RAW) cultured alone or co-cultured under the suppression of de novo ceramide synthesis. Palmitate enhanced ceramide accumulation and stimulated the expression and secretion of interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) in L1. The suppression of serine-palmitoyl transferase, a rate-limiting enzyme of de novo ceramide synthesis, by myriocin or siRNA attenuated those palmitate-induced alterations, and a ceramide synthase inhibitor fumonisin B1 showed similar results. In contrast, the inhibitor of sphingosine kinase or a membrane-permeable ceramide analogue augmented the cytokine secretion. Myriocin effects on the palmitate-induced changes were not abrogated by toll-like receptor-4 blockade. Although palmitate stimulated RAW to secrete tumor necrosis factor-alpha (TNF-alpha), it did not significantly increase ceramide content, and neither myriocin nor fumonisin B1 attenuated the TNF-alpha hypersecretion. The co-culture of L1 with RAW markedly augmented IL-6 and MCP-1 levels in media. Myriocin or fumonisin B1 significantly lowered these cytokine levels and suppressed the gene expression of TNF-alpha and MCP-1 in RAW and of IL-6 and MCP-1 in L1. In conclusion, de novo synthesized ceramide partially mediates the palmitate effects on pro-inflammatory adipokines and is possibly involved in the interaction with macrophages. CI - Copyright (c) 2014 Elsevier Inc. All rights reserved. FAU - Hamada, Yoji AU - Hamada Y AD - Department of Metabolic Medicine, Nagoya University Graduate School of Medicine, 65 Tsuruma-cho, Showa-ku, Nagoya 466-8550, Japan. Electronic address: yhama@med.nagoya-u.ac.jp. FAU - Nagasaki, Hiroshi AU - Nagasaki H AD - Department of Metabolic Medicine, Nagoya University Graduate School of Medicine, 65 Tsuruma-cho, Showa-ku, Nagoya 466-8550, Japan; Department of Physiology I, School of Medicine, Fujita Health University, 1-98 Kutsukake-cho, Toyoake, Aichi 470-1192, Japan. FAU - Fujiya, Atsushi AU - Fujiya A AD - Department of Endocrinology and Diabetes, Nagoya University Graduate School of Medicine, 65 Tsuruma-cho, Showa-ku, Nagoya 466-8550, Japan. FAU - Seino, Yusuke AU - Seino Y AD - Department of Metabolic Medicine, Nagoya University Graduate School of Medicine, 65 Tsuruma-cho, Showa-ku, Nagoya 466-8550, Japan. FAU - Shang, Qing-Long AU - Shang QL AD - Pharmaceutical Research Laboratories, Sanwa Kagaku Kenkyusho Co, Ltd, 363 Shiosaki, Hokusei-cho, Inabe 511-0406, Japan; Department of Microbiology, Harbin Medical University, Harbin 150086, China. FAU - Suzuki, Takeshi AU - Suzuki T AD - Pharmaceutical Research Laboratories, Sanwa Kagaku Kenkyusho Co, Ltd, 363 Shiosaki, Hokusei-cho, Inabe 511-0406, Japan. FAU - Hashimoto, Hiroyuki AU - Hashimoto H AD - Pharmaceutical Research Laboratories, Sanwa Kagaku Kenkyusho Co, Ltd, 363 Shiosaki, Hokusei-cho, Inabe 511-0406, Japan. FAU - Oiso, Yutaka AU - Oiso Y AD - Department of Endocrinology and Diabetes, Nagoya University Graduate School of Medicine, 65 Tsuruma-cho, Showa-ku, Nagoya 466-8550, Japan. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20140919 PL - United States TA - J Nutr Biochem JT - The Journal of nutritional biochemistry JID - 9010081 RN - 0 (Adipokines) RN - 0 (Ccl2 protein, mouse) RN - 0 (Ceramides) RN - 0 (Chemokine CCL2) RN - 0 (Fatty Acids, Monounsaturated) RN - 0 (Fumonisins) RN - 0 (Interleukin-6) RN - 0 (Palmitates) RN - 0 (RNA, Messenger) RN - 0 (Tlr4 protein, mouse) RN - 0 (Toll-Like Receptor 4) RN - 0 (Tumor Necrosis Factor-alpha) RN - 3ZZM97XZ32 (fumonisin B1) RN - EC 2.3.1.50 (Serine C-Palmitoyltransferase) RN - YRM4E8R9ST (thermozymocidin) SB - IM MH - 3T3-L1 Cells MH - Adipocytes/*metabolism MH - Adipokines/*metabolism MH - Animals MH - Cell Line, Tumor MH - Ceramides/*biosynthesis MH - Chemokine CCL2/genetics/metabolism MH - Coculture Techniques MH - Fatty Acids, Monounsaturated/pharmacology MH - Fumonisins/pharmacology MH - Interleukin-6/genetics/metabolism MH - Macrophages/*metabolism MH - Mice MH - Palmitates/pharmacology MH - RNA, Messenger/genetics/metabolism MH - Serine C-Palmitoyltransferase/metabolism MH - Toll-Like Receptor 4/genetics/metabolism MH - Tumor Necrosis Factor-alpha/genetics/metabolism OTO - NOTNLM OT - Adipocytes OT - Ceramide OT - Cytokines OT - Macrophages OT - Palmitate OT - Serine-palmitoyl transferase EDAT- 2014/10/07 06:00 MHDA- 2015/08/08 06:00 CRDT- 2014/10/07 06:00 PHST- 2014/01/15 00:00 [received] PHST- 2014/05/14 00:00 [revised] PHST- 2014/07/24 00:00 [accepted] PHST- 2014/10/07 06:00 [entrez] PHST- 2014/10/07 06:00 [pubmed] PHST- 2015/08/08 06:00 [medline] AID - S0955-2863(14)00172-7 [pii] AID - 10.1016/j.jnutbio.2014.07.008 [doi] PST - ppublish SO - J Nutr Biochem. 2014 Dec;25(12):1309-16. doi: 10.1016/j.jnutbio.2014.07.008. Epub 2014 Sep 19.