Release of a specific set of proinflammatory adipokines by differentiating 3T3-L1 cells

Objective: Although there is a large amount of data on the role of preadipocytes in promoting the release of proinflammatory cytokines and chemokines in response to macrophage-derived cytokines, the direct role of insulin and saturated fatty acids in modulating the release of inflammatory cytokines by cells differentiating along the adipocytic lineage is less understood. Methods: 3T3-L1 murine preadipocyte cells were cultured for 3 d in a proliferating medium in the presence or absence of insulin 0.2 nmol/L plus palmitic acid 1 mu mol/L. In parallel, 3T3-L1 cells were cultured in a differentiation medium containing dexamethasone, insulin, and isobutyl methyl xanthine in the absence or presence of palmitic acid for 3 d. The levels of several cytokines were evaluated in the culture supernatants by a bead-based multiplex immunoassay. Results: Under the proliferation conditions, insulin plus palmitic acid promoted a significant increase in the release of interleukin-6, keratinocyte-derived chemokine (KC)/chemokine ligand-1 (CXCL-1), monocyte chemotactic protein-1 (MCP-1), and regulated and normal T cells expressed and presumably secreted (RANTES) by 31341 preadipocytes. Of note, the release of KC/CXCL-1, MCP-1, and RANTES increased significantly with adipocytic differentiation, and the addition of palmitic acid to differentiating 3T3-L1 cells resulted in a further significant promotion of KC/CXCL1, MCP-1, and RANTES, coupled to the increase of additional cytokines. Conclusions: Taken together, these data show that a restricted common group of cytokines/chemokines (KC/CXCL1. MCP-1, and RANTES) is upregulated in proliferating and differentiating 313-L1 adipocytes in response to insulin and palmitic acid and that differentiating adipocytes respond with an increased range of cytokines with respect to proliferating 3T3-L1 preadipocytes. (C) 2013 Elsevier Inc. All rights reserved.