Romacho Tania, Glosse Philipp, Richter Isabel, Elsen Manuela, Schoemaker Marieke H, van Tol Eric A, Eckel Jürgen
Paul Langerhans Group for Integrative Physiology, German Diabetes Center, Auf'm Hennekamp 65, 40225 Düsseldorf, Germany.
Global Discovery, Mead Johnson Pediatric Nutrition Institute, Middenkampweg 2, Nijmegen, 6545 CJ, The Netherlands.
Nutrients. 2015 Jan 26;7(2):865-86. doi: 10.3390/nu7020865.
Nutritional factors such as casein hydrolysates and long chain polyunsaturated fatty acids have been proposed to exert beneficial metabolic effects. We aimed to investigate how a casein hydrolysate (eCH) and long chain polyunsaturated fatty acids could affect human primary adipocyte function in vitro. Incubation conditions with the different nutritional factors were validated by assessing cell vitality with lactate dehydrogenase (LDH) release and neutral red incorporation. Intracellular triglyceride content was assessed with Oil Red O staining. The effect of eCH, a non-peptidic amino acid mixture (AA), and long-chain polyunsaturated fatty acids (LC-PUFAs) on adiponectin and leptin secretion was determined by enzyme-linked immunosorbent assay (ELISA). Intracellular adiponectin expression and nuclear factor-κB (NF-κB) activation were analyzed by Western blot, while monocyte chemoattractant protein-1 (MCP-1) release was explored by ELISA. The eCH concentration dependently increased adiponectin secretion in human primary adipocytes through its intrinsic peptide bioactivity, since the non-peptidic mixture, AA, could not mimic eCH's effects on adiponectin secretion. Eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and DHA combined with arachidonic acid (ARA) upregulated adiponectin secretion. However, only DHA and DHA/ARA exerted a potentanti-inflammatory effect reflected by prevention of tumor necrosis factor-α (TNF-α) induced NF-κB activation and MCP-1 secretion in human adipocytes. eCH and DHA alone or in combination with ARA, may hold the key for nutritional programming through their anti-inflammatory action to prevent diseases with low-grade chronic inflammation such as obesity or diabetes.
酪蛋白水解物和长链多不饱和脂肪酸等营养因素已被提出具有有益的代谢作用。我们旨在研究酪蛋白水解物(eCH)和长链多不饱和脂肪酸如何在体外影响人原代脂肪细胞功能。通过用乳酸脱氢酶(LDH)释放和中性红摄取评估细胞活力来验证不同营养因素的孵育条件。用 Oil Red O 染色评估细胞内甘油三酯含量。通过酶联免疫吸附测定(ELISA)测定 eCH、非肽氨基酸混合物(AA)和长链多不饱和脂肪酸(LC-PUFA)对脂联素和瘦素分泌的影响。通过蛋白质免疫印迹分析细胞内脂联素表达和核因子-κB(NF-κB)激活,同时通过 ELISA 探索单核细胞趋化蛋白-1(MCP-1)释放。eCH 通过其内在的肽生物活性浓度依赖性地增加人原代脂肪细胞中脂联素的分泌,因为非肽混合物 AA 不能模拟 eCH 对脂联素分泌的影响。二十碳五烯酸(EPA)、二十二碳六烯酸(DHA)以及 DHA 与花生四烯酸(ARA)的组合上调了脂联素的分泌。然而,只有 DHA 和 DHA/ARA 发挥了强大的抗炎作用,表现为在人脂肪细胞中预防肿瘤坏死因子-α(TNF-α)诱导的 NF-κB 激活和 MCP-1 分泌。单独的 eCH 和 DHA 或与 ARA 联合使用,可能通过其抗炎作用成为营养编程的关键,以预防肥胖或糖尿病等低度慢性炎症疾病。