Pepsin-pancreatin protein hydrolysates from extruded amaranth inhibit markers of atherosclerosis in LPS-induced THP-1 macrophages-like human cells by reducing expression of proteins in LOX-1 signaling pathway

Abstract

BackgroundAtherosclerosis is considered a progressive disease that affects arteries that bring blood to the heart, to the brain and to the lower end. It derives from endothelial dysfunction and inflammation, which play an important role in the thrombotic complications of atherosclerosis. Cardiovascular disease is the leading cause of death around the world and one factor that can contribute to its progression and prevention is diet. Our previous study found that amaranth hydrolysates inhibited LPS-induced inflammation in human and mouse macrophages by preventing activation of NF-κB signaling. Furthermore, extrusion improved the anti-inflammatory effect of amaranth protein hydrolysates in both cell lines, probably attributed to the production of bioactive peptides during processing. Therefore, the objective of this study was to compare the anti-atherosclerotic potential of pepsin-pancreatin hydrolysates from unprocessed and extruded amaranth in THP-1 lipopolysaccharide-induced human macrophages and suggest the mechanism of action.ResultsUnprocessed amaranth hydrolysate (UAH) and extruded amaranth hydrolysate (EAH) showed a significant reduction in the expression of interleukin-4 (IL-4) (69% and 100%, respectively), interleukin-6 (IL-6) (64% and 52%, respectively), interleukin-22 (IL-22) (55% and 70%, respectively). Likewise, UAH and EAH showed a reduction in the expression of monocyte-chemo attractant protein-1 (MCP-1) (35% and 42%, respectively), transferrin receptor-1 (TfR-1) (48% and 61%, respectively), granulocyte-macrophage colony-stimulating factor (GM-CSF) (59% and 63%, respectively), and tumor necrosis factor-α (TNF-α) (60% and 63%, respectively). Also, EAH reduced the expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) (27%), intracellular adhesion molecule-1 (ICAM-1) (28%) and matrix metalloproteinase-9 (MMP-9) (19%), important molecular markers in the atherosclerosis pathway. EAH, led to a reduction of 58, 52 and 79% for LOX-1, ICAM-1 and MMP-9, respectively, by confocal microscopy.ConclusionsExtruded amaranth hydrolysate showed potential anti-atherosclerotic effect in LPS-induced THP-1 human macrophage-like cells by reducing the expression of proteins associated with LOX-1 signaling pathway.

Intake of Protein Hydrolysates and Phenolic Fractions Isolated from Flaxseed Ameliorates TNBS‐Induced Colitis

“…Upon cell stimulation, IκB proteins are phosphorylated by IκB kinases, releasing the NF‐κB, which is then translocated to the nucleus, inducing the expression of various genes . It has been recently shown that peptides derived from some foods are capable of promoting the inactivation of the IkB kinase complex or inhibiting the translocation of p50 and p65 subunits of NF‐κB . Thus, we also suggest that the capability of flaxseed peptides to inhibit pro‐inflammatory mediators may occur through the inactivation of IκB kinase complex or by the inhibition of the translocation of NF‐κB p50 and p65 subunits.…”

“…[36] It has been recently shown that peptides derived from some foods are capable of promoting the inactivation of the IkB kinase complex or inhibiting the translocation of p50 and p65 subunits of NF-κB. [35,41,42] Thus, we also suggest that the capability of flaxseed peptides to inhibit pro-inflammatory mediators may occur through the inactivation of IκB kinase complex or by the inhibition of the translocation of NF-κB p50 and p65 subunits. Further experiments are necessary to prove our hypothesis.…”

Chia (Salvia hispanica L.) Seed Total Protein and Protein Fractions Digests Reduce Biomarkers of Inflammation and Atherosclerosis in Macrophages In Vitro

“…These monocytes differentiated into macrophages and phagocyte ox‐LDL. We observed a reduction of expression of ICAM and LOX‐1 by macrophages treated with DTP and DPF, resulting in less formation of foam cells, similar with unprocessed and extruded amaranth hydrolysate …”

“…We observed a reduction of expression of ICAM and LOX-1 by macrophages treated with DTP and DPF, resulting in less formation of foam cells, similar with unprocessed and extruded amaranth hydrolysate. [59] These foam cells secrete MCP-1, a chemokine that binds to the CCR2 and attracts new monocytes, increasing the atherosclerosis plaque. [4] Also, the accumulation of modified LDL by macrophages activates cytokine production, like TNF-α, IL-6, IL-1β, and IL-12, and active the enzymes iNOS and COX-2 to Figure 9.…”

Food‐derived immunomodulatory peptides

“…Another impact of processing was evaluated during in vitro digestion of unprocessed and extruded amaranth flour with hydrolysates, where the time to hydrolysis was considered. The results showed that enzymatic (pepsin and pancreatin) hydrolysis and a pre‐treatment of the extrusion (125 °C and screw speed of 130 rpm) promoted the released of peptides of low molecular mass after 10 min of hydrolysis, while those from unprocessed amaranth flour were released after 25 and 60 min of hydrolysis . This could be due to the exposure of food proteins to high pressure, temperature and shear forces for a short amount of time, which could denature proteins and make them more available to enzymatic action …”