Monk Jennifer M, Liddle Danyelle M, De Boer Anna A, Brown Morgan J, Power Krista A, Ma David Wl, Robinson Lindsay E
Department of Human Health and Nutritional Sciences, University of Guelph, Guelph, Canada; and Guelph Food Research Centre, Agriculture Agri-Food Canada, Guelph, Canada.
Department of Human Health and Nutritional Sciences, University of Guelph, Guelph, Canada; and.
J Nutr. 2015 Apr;145(4):829-38. doi: 10.3945/jn.114.205443. Epub 2015 Feb 4.
Obese adipose tissue (AT) inflammation is characterized by dysregulated adipokine production and immune cell accumulation. Cluster of differentiation (CD) 8+ T cell AT infiltration represents a critical step that precedes macrophage infiltration. n-3 (ω-3) Polyunsaturated fatty acids (PUFAs) exert anti-inflammatory effects in obese AT, thereby disrupting AT inflammatory paracrine signaling.
We assessed the effect of n-3 PUFAs on paracrine interactions between adipocytes and primary CD8+ T cells co-cultured at the cellular ratio observed in obese AT.
C57BL/6 mice were fed either a 3% menhaden fish-oil + 7% safflower oil (FO) diet (wt:wt) or an isocaloric 10% safflower oil (wt:wt) control (CON) for 3 wk, and splenic CD8+ T cells were isolated by positive selection (via magnetic microbeads) and co-cultured with 3T3-L1 adipocytes. Co-cultures were unstimulated (cells alone), T cell receptor stimulated, or lipopolysaccharide (LPS) stimulated for 24 h.
In LPS-stimulated co-cultures, FO reduced secreted protein concentrations of interleukin (IL)-6 (-42.6%), tumor necrosis factor α (-67%), macrophage inflammatory protein (MIP) 1α (-52%), MIP-1β (-62%), monocyte chemotactic protein (MCP) 1 (-23%), and MCP-3 (-19%) vs. CON, which coincided with a 74% reduction in macrophage chemotaxis toward secreted chemotaxins in LPS-stimulated FO-enriched co-culture-conditioned media. FO increased mRNA expression of the inflammatory signaling negative regulators monocyte chemoattractant 1-induced protein (Mcpip; +9.3-fold) and suppressor of cytokine signaling 3 (Socs3; +1.7-fold), whereas FO reduced activation of inflammatory transcription factors nuclear transcription factor κB (NF-κB) p65 and signal transducer and activator of transcription 3 (STAT3) by 27% and 33%, respectively. Finally, mRNA expression of the inflammasome components Caspase1 (-36.4%), Nod-like receptor family pyrin domain containing 3 (Nlrp3; -99%), and Il1b (-68.8%) were decreased by FO compared with CON (P ≤ 0.05).
FO exerted an anti-inflammatory and antichemotactic effect on the cross-talk between CD8+ T cells and adipocytes and has implications in mitigating macrophage-centered AT-driven components of the obese phenotype.
肥胖脂肪组织（AT）炎症的特征是脂肪因子产生失调和免疫细胞聚集。分化簇（CD）8+T细胞浸润到AT中是巨噬细胞浸润之前的关键步骤。n-3（ω-3）多不饱和脂肪酸（PUFAs）在肥胖的AT中发挥抗炎作用，从而破坏AT炎症旁分泌信号。
我们评估了n-3 PUFAs对在肥胖AT中观察到的细胞比例下共培养的脂肪细胞与原代CD8+T细胞之间旁分泌相互作用的影响。
将C57BL/6小鼠喂食3%的鲱鱼油+7%的红花油（FO）饮食（重量比）或等热量的10%红花油（重量比）对照饮食（CON）3周，通过阳性选择（通过磁性微珠）分离脾CD8+T细胞，并与3T3-L1脂肪细胞共培养。共培养物在未刺激（仅细胞）、T细胞受体刺激或脂多糖（LPS）刺激下培养24小时。
在LPS刺激的共培养物中，与CON相比，FO降低了白细胞介素（IL）-6（-42.6%）、肿瘤坏死因子α（-67%）、巨噬细胞炎性蛋白（MIP）1α（-52%）、MIP-1β（-62%）、单核细胞趋化蛋白（MCP）1（-23%）和MCP-3（-19%）的分泌蛋白浓度，这与LPS刺激的富含FO的共培养条件培养基中巨噬细胞对分泌趋化因子的趋化作用降低74%一致。FO增加了炎症信号负调节因子单核细胞趋化因子1诱导蛋白（Mcpip；+9.3倍）和细胞因子信号抑制因子3（Socs3；+1.7倍）的mRNA表达，而FO分别将炎症转录因子核转录因子κB（NF-κB）p65和信号转导子及转录激活子3（STAT3）的激活降低了27%和33%。最后，与CON相比，FO降低了炎性小体成分半胱天冬酶1（-36.4%）、含NOD样受体家族吡咯结构域3（Nlrp3；-99%）和Il1b（-68.8%）的mRNA表达（P≤0.05）。
FO对CD8+T细胞与脂肪细胞之间的相互作用具有抗炎和抗趋化作用，并对减轻肥胖表型中以巨噬细胞为中心的AT驱动成分具有重要意义。
