Abstract
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.
Highlights
The prevalence of childhood obesity has dramatically increased in recent years [1]
In order to validate the incubation conditions in our cell system, we explored the effect of the casein hydrolysate extensive casein hydrolysates (eCH) and a non-peptide amino acid mixture (AA) on parameters of cell cytotoxicity such as lactate dehydrogenase (LDH, L-lactate:NAD oxidoreductase, EC. 1.1.1.27)
In order to further determine if this potent anti-inflammatory effect exerted by docosahexaenoic acid (DHA) alone or with arachidonic acid (ARA) translated to a reduction of downstream targets of nuclear factor-κB (NF-κB), we studied the effect of the pre-incubation with Eicosapentaenoic acid (EPA), DHA, ARA and the DHA/ARA
Summary
It has been proposed that rapid weight gain during the first years of life is related to the development of obesity later in life [2]. The impact of nutritional factors on the development of obesity and its related metabolic complications later in adult life has generated considerable interest in the field of infant nutrition [5,6]. AT enlargement triggers an imbalance in adipokine secretion. In these diseases the circulating levels of several pro-inflammatory adipokines, such as leptin and TNF-α, are enhanced, followed by a lower expression of adiponectin, which contributes to the characteristic low-grade chronic inflammatory status [8,9,10]. AT inflammation represents a potential key pathogenic link between obesity, insulin resistance, and type 2 diabetes [7,11]
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