Palmitic acid triggers inflammatory responses in N42 cultured hypothalamic cells partially via ceramide synthesis but not via TLR4

reserved10siFinanziato da Scottish Universities Life Sciences Alliance (SULSA-MSD) studentship, the University of Aberdeen and the Scottish Government's Rural e Environment Science and Analytical Services Division (RESAS).A high-fat diet induces hypothalamic inflammation in rodents which, in turn, contributes to the development of obesity by eliciting both insulin and leptin resistance. However, the mechanism by which long-chain saturated fatty acids trigger inflammation is still contentious. To elucidate this mechanism, the effect of fatty acids on the expression of the pro-inflammatory cytokinesIL-6andTNF alpha was investigated in the mHypoE-N42 hypothalamic cell line (N42). N42 cells were treated with lauric acid (LA) and palmitic acid (PA). PA challenge was carried out in the presence of either a TLR4 inhibitor, a ceramide synthesis inhibitor (L-cycloserine), oleic acid (OA) or eicosapentaenoic acid (EPA). Intracellular ceramide accumulation was quantified using LC-ESI-MS/MS. PA but not LA upregulatedIL-6andTNF alpha. L-cycloserine, OA and EPA all counteracted PA-induced intracellular ceramide accumulation leading to a downregulation ofIL-6andTNF alpha. However, a TLR4 inhibitor failed to inhibit PA-induced upregulation of pro-inflammatory cytokines. In conclusion, PA induced the expression of IL-6 and TNF alpha in N42 neuronal cells independently of TLR4 but, partially, via ceramide synthesis with OA and EPA being anti-inflammatory by decreasing PA-induced intracellular ceramide build-up. Thus, ceramide accumulation represents one on the mechanisms by which PA induces inflammation in neurons.mixedSergi, Domenico; Morris, Amanda C; Kahn, Darcy E; McLean, Fiona H; Hay, Elizabeth A; Kubitz, Phil; MacKenzie, Alasdair; Martinoli, Maria G; Drew, Janice E; Williams, Lynda MSergi, Domenico; Morris, Amanda C; Kahn, Darcy E; Mclean, Fiona H; Hay, Elizabeth A; Kubitz, Phil; Mackenzie, Alasdair; Martinoli, Maria G; Drew, Janice E; Williams, Lynda