casein-hydrolysate and Inflammation

Topics: 3T3-L1 Cells; Animals; Caseins; Cattle; Diet, High-Fat; Inflammation; Insulin Resistance; Mice; Mice, Inbred C57BL; Mice, Obese; Mitogen-Activated Protein Kinases; Obesity; Tumor Necrosis Factor-alpha

During the thermal processing of dairy products, the Maillard reaction occurs between milk proteins and lactose, resulting in the formation of various products including glycated proteins. In this study, lactose-glycated casein was generated through the Maillard reaction between casein and lactose and then hydrolyzed by a trypsin preparation. The anti-inflammatory effect of the resultant glycated casein hydrolysate (GCH) was investigated using the lipopolysaccharide (LPS)-sitmulated rat intestinal epithelial (IEC-6) cells as a cell model and corresponding casein hydrolysate (CH) as a control. The results indicated that the preformed glycation enabled lactose conjugation to casein, which endowed GCH with a lactose content of 12.61 g/kg protein together with a lower activity than CH to enhance the viability value of the IEC-6 cells. The cells with LPS stimulation showed significant inflammatory responses, while a pre-treatment of the cells with GCH before LPS stimulation consistently led to a decreased secretion of three pro-inflammatory mediators, namely, IL-6, IL-1β and tumor necrosis factor-α (TNF-α) but an increased secretion of two anti-inflammatory mediators, including IL-10 and transforming growth factor-β (TGF-β), demonstrating the anti-inflammatory potential of GCH in LPS-stimulated cells. In addition, GCH up-regulated the expression of TLR4, p-p38, and p-p65 proteins in the stimulated cells, resulting in the suppression of NF-κB and MAPK signaling pathways. Collectively, GCH was mostly less efficient than CH to exert these assessed anti-inflammatory activities in the cells and more importantly, GCH also showed an ability to cause cell inflammation by promoting IL-6 secretion and up-regulating the expression of TLR4 and p-p65. The casein lactose-glycation of the Maillard-type was thereby concluded to attenuate the anti-inflammatory potential of the resultant casein hydrolysate. It is highlighted that the casein lactose-glycation of the Maillard-type might cause a negative impact on the bioactivity of casein in the intestine, because the glycated casein after digestion could release GCH with reduced anti-inflammatory activity.

Topics: Animals; Anti-Inflammatory Agents; Caseins; Inflammation; Interleukin-6; Lactose; Lipopolysaccharides; Maillard Reaction; NF-kappa B; Rats

Modulation of inflammation-related immune response on THP-1 macrophages of protein hydrolysates derived from tilapia mince, casein and pea protein, were investigated. The protein substrates were hydrolyzed by Virgibacillus halodenitrificans SK1-3-7 proteinase. The degree of hydrolysis (DH) of casein was observed to be the highest throughout the course of hydrolysis. When challenging THP-1 macrophages, tilapia mince hydrolysate (TMH) enhanced innate immunity through induction of IL-1β and COX-2 expression. Anti-inflammatory activity was observed in casein hydrolysate (CH) and pea protein hydrolysate (PPH) by attenuating lipopolysaccharide- (LPS) induced pro-inflammatory gene expression in THP-1 macrophages. CH suppressed IL-1β, IL-6, IL-8, TNF-α and COX-2, while PPH reduced LPS-induced IL-6 and TNF-α responses. In addition, CH and PPH showed stronger suppression of LPS-induced pro-inflammatory gene expression compared with non-hydrolyzed casein and pea protein. These results suggest that TMH, CH and PPH prepared from V. halodenitrificans SK1-3-7 proteinase are potential functional food ingredients with immunomodulatory activity.

Topics: Animals; Anti-Inflammatory Agents; Caseins; Cells, Cultured; Cichlids; Cyclooxygenase 2; Fish Proteins; Gene Expression Regulation; Humans; Hydrolysis; Immunity, Innate; Immunomodulation; Inflammation; Interleukin-1beta; Interleukin-6; Interleukin-8; Lipopolysaccharides; Macrophages; Peptide Hydrolases; Pisum sativum; Plant Proteins; Protein Hydrolysates; Tumor Necrosis Factor-alpha; Virgibacillus

Activation of the nod-like receptor protein 3 (NLRP3) inflammasome is required for IL-1β release and is a key component of obesity-induced inflammation and insulin resistance. This study hypothesized that supplementation with a casein hydrolysate (CH) would attenuate NLRP3 inflammasome mediated IL-1β secretion in adipose tissue (AT) and improve obesity-induced insulin resistance.. J774.2 macrophages were LPS primed (10 ng/mL) and stimulated with adenosine triphosphate (5 mM) to assess NLRP3 inflammasome activity. Pretreatment with CH (1 mg/mL; 48 h) reduced caspase-1 activity and decreased IL-1β secretion from J774.2 macrophages in vitro. 3T3-L1 adipocytes cultured with conditioned media from CH-pretreated J774.2 macrophages demonstrated increased phosphorylated (p)AKT expression and improved insulin sensitivity. C57BL/6JOLaHsd mice were fed chow or high fat diet (HFD) for 12 wk ± CH resuspended in water (0.5% w/v). CH supplementation improved glucose tolerance in HFD-fed mice as determined by glucose tolerance test. CH supplementation increased insulin-stimulated pAKT protein levels in AT, liver, and muscle after HFD. Cytokine secretion was measured from AT and isolated bone marrow macrophages cultured ex vivo. CH supplementation attenuated IL-1β, tumor necrosis factor alpha (TNF-α) and IL-6 secretion from AT and IL-1β, IL-18, and TNF-α from bone marrow macrophages following adenosine triphosphate stimulation ex vivo.. This novel CH partially protects mice against obesity-induced hyperglycemia coincident with attenuated IL-1β secretion and improved insulin signaling.

Topics: 3T3-L1 Cells; Adipose Tissue; Animals; Caseins; Cytokines; Diabetes Mellitus, Type 2; Diet, High-Fat; Hyperglycemia; Inflammasomes; Inflammation; Insulin; Insulin Resistance; Interleukin-1beta; Interleukin-6; Mice; Mice, Inbred C57BL; NLR Family, Pyrin Domain-Containing 3 Protein; NLR Proteins; Obesity; Tumor Necrosis Factor-alpha

This study was designed to determine whether marine-derived proteins other than cod could have beneficial effects on inflammation following muscle injury. Macrophage and neutrophil densities were measured from bupivacaine-injured tibialis anterior muscle of rats fed isoenergetic diets containing either shrimp hydrolysate (Shr), casein hydrolysate (CaH), or whole casein (Ca). In this study, Shr reduced ED

Topics: Animals; Bupivacaine; Caseins; Inflammation; Macrophages; Muscle, Skeletal; Pandalidae; Protein Hydrolysates; Rats; Wounds and Injuries

Bioactive peptides from milk can impart a wide range of physiological benefits without the allergies and intolerance associated with the consumption of whole milk. The objective of this study was to characterise the anti-inflammatory properties of intact sodium caseinate (NaCAS), a moderately hydrolysed NaCAS enzyme hydrolysate (EH) and its 5 kDa fraction (5kDaR), in both in vitro and ex vivo systems. In vitro, Caco-2 cells were stimulated with tumor necrosis factor (TNF) α and co-treated ± casein hydrolysates or dexamethasone (control). The inflammatory marker interleukin (IL)-8 was measured by ELISA in the supernatant at 24 h. Ex vivo, porcine colonic tissues were stimulated with lipopolysaccharide (LPS) and co-treated with casein hydrolysates for 3 h from which the relative expression of a panel of cytokines was measured in vitro. While the steroid dexamethasone brought about a 41.6% reduction in the IL-8 concentration in the supernatant, the 5kDaR reduced IL-8 by 59% (P < 0.05) when compared to the TNFα stimulated Caco-2 cells. In the ex vivo system, 5kDaR was associated with decreases in IL-1α, IL-1β, IL-8 and TGF-β expression and an increase in IL-17 expression (P < 0.05) relative to the LPS challenged tissues. We concluded, that a 5 kDa casein fraction demonstrates potent anti-inflammatory effects both in in vitro and ex vivo models of the gastrointestinal tract.

Topics: Animals; Anti-Inflammatory Agents; Caco-2 Cells; Caseins; Dexamethasone; Gastrointestinal Tract; Humans; Inflammation; Interleukin-17; Interleukin-1alpha; Interleukin-1beta; Interleukin-8; Lipopolysaccharides; Models, Biological; Molecular Weight; Swine; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

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.

Topics: Adipocytes; Adipokines; Caseins; Cell Culture Techniques; Chemokine CCL2; Docosahexaenoic Acids; Eicosapentaenoic Acid; Fatty Acids, Unsaturated; Humans; Inflammation; L-Lactate Dehydrogenase; Leptin; Milk Proteins; NF-kappa B; Tumor Necrosis Factor-alpha

Nonsteroidal anti-inflammatory drugs (NSAID) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue. Earlier studies in our laboratory have found that specific casein hydrolysates (CH) might be useful in the treatment of gastrointestinal wounds. The underlying mechanisms that support inflammation and wound healing are not completely understood, but transcriptional alterations may be used as markers for inflammation and wound healing. The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin (60 min) followed by corolase (0, 10, or 60 min) were investigated in intestinal epithelial cells treated with the NSAID indomethacin. The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 (TGF-β1), cyclooxygenase-2 (COX-2), peroxisome proliferator-activated receptor-γ (PPAR-γ) and nuclear factor κB (NFκB) by real-time PCR. Furthermore, the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels. Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB (P < 0.05) compared with the hydrolysate treated with pepsin only. Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of COX-2 (P < 0.05) compared with hydrolysate treated with corolase for only 10 min whereas transcription of PPAR-γ was not affected (P > 0.05). Additionally, the hydrolysate prepared by pepsin treatment only (0 min corolase) had a pro-inflammatory effect on macrophages via PG E(2) stimulation (P < 0.05). In conclusion, CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1, COX-2, and NFκB.

Topics: Animals; Biomarkers; Caseins; Cell Line; Cytokines; Epithelial Cells; Gene Expression Regulation; Inflammation; Intestinal Mucosa; Macrophages; Mice; Models, Biological; Rats; Real-Time Polymerase Chain Reaction

Mouse experiments were conducted in order to find whether oral application of tryptic casein hydrolysate (TCH) results in enhancement of phagocytosing capacity of murine phagocytic cells as well as whether such application might be of use for prevention of inflammatory processes. Phagocytosing capacity of phagocytic cells of mice that received oral TCH once daily in a dose of 1.0 mg/g body weight dissolved in 0.5 ml of distilled water for five successive days was significantly higher (p < 0.05) than that of mice given equivalent volumes of distilled water, with a phagocytosing capacity enhancement index being 1.39 and 1.34 regarding peritoneal macrophages and blood phagocytic cells, respectively. Taken on the other hand, the immunostimulatory effects of oral TCH were found to be not enough to prevent mice from inflammation that was induced experimentally using acute (paw edema) and contact hypersensitivity models. A possibility for development of food protein enzymatic hydrolysates as antimicrobial immunostimulants acting through improvement of phagocytic cell functioning is discussed.

Topics: Administration, Oral; Animals; Carrageenan; Caseins; Dermatitis, Contact; Dinitrofluorobenzene; Disease Models, Animal; Edema; Female; Granulocytes; Inflammation; Interleukin-10; Macrophages, Peritoneal; Male; Mice; Mice, Inbred BALB C; Monocytes; Phagocytes; Phagocytosis; Trypsin; Tumor Necrosis Factor-alpha