GPETAFLR, a novel bioactive peptide from Lupinus angustifolius L. protein hydrolysate, reduces osteoclastogenesis

Abstract

26The effect of GPETAFLR, a peptide isolated from Lupinus angustifolius L. 27 protein hydrolysate (LPH), on osteoclastogenesis was investigated. Human 28 osteoclasts generated from monocytes were used to analyse the effects of 29 GPETAFLR (50-100 µg/mL) on osteoclastogenesis using TRAP reaction, RT-30 qPCR, and ELISA procedures. LPS enhanced TRAP activity and the expression 31 of osteoclast marker genes (TRAP, OSCAR, RANK, and CATHK) while 32 downregulated the expression of OPG gene in human monocyte-derived 33 osteoclasts. These effects were reduced with GPETAFLR. Moreover, LPS 34 increased the release of osteoclastogenic cytokines (TNF-, IL-1, and IL-6) 35 meanwhile GPETAFLR increased the release of anti-osteoclastogenic cytokines 36 (IL-4 and IL-10) in the medium of human monocyte-derived osteoclasts. For the 37 first time, we show that plant peptides from lupine protein hydrolysates have 38 anti-osteoclastogenic activity. These exciting findings open opportunities for 39 developing nutritional strategies with Lupinus angustifolius L. as dietary source 40 of plant proteins, notably GPETAFLR, to prevent development and progression 41 of osteoclast-related diseases. 42 43 44 45 46 47 65 cells that adhere to the bone matrix and secrete acids and lithic enzymes 66 resulting in bone degradation (Kalbasi Anaraki et al., 2015). The differentiation 67 of monocytes into OCs, a multi-complex procedure referred to as 68 osteoclastogenesis, is induced by two crucial cytokines: macrophage colony-69 stimulating factor (M-CSF) and RANKL. In addition to M-CSF and RANKL as 70 the only factors required for basal osteoclastogenesis, several other cytokines 71 such as TNF-, IL-1, and IL-6 also play an important role in OC biology. These 72 osteoclastogenic cytokines promote bone resorption through multiple 73 mechanisms that include the increase of M-CSF and RANKL production 74 (Zupan, Jeras, & Marc, 2013). 307 T.M., … Oliveira, R.C. (2014). Effects of fluoride in bone repair: an evaluation of 308 RANKL, OPG and TRAP expression. Odontology, 102, 22-30.

GPETAFLR, a biopeptide from Lupinus angustifolius L., protects against oxidative and inflammatory damage in retinal pigment epithelium cells

“…Researchers found that buckwheat extracts may inhibit adipogenesis and inflammatory response during adipocyte differentiation of 3T3‐L1 cells.22 Brazilian red propolis ( Apis mellifera ), Copaifera oleoresins, flavonoid fraction of Bergamot Juice ( Citrus bergamia ), effusanin C ( Isodon japonicus ), oligomeric proanthocyanidins ( crataegus oxyacantha ) are others isolated compounds with anti‐inflammatory actions in activated monocytes and macrophages (Montserrat‐de la Paz et al, ). As well as prior studies (Millan‐Linares et al, , ), showing GPETAFLR as one of the major bioactive peptides with anti‐inflammatory activity, our observations suggest that this octapeptide isolated from Lupinus angustifolius L . is associated with a remarkable prevention of oxidative and inflammatory in RPE cells.…”

“…Considering the MTT assay, 50 and 100 μg/ml of peptide showed more than 98% viability of the cells. In previous reports, GPETAFLR were not cytotoxic to primary human monocytes (Montserrat‐de la Paz et al, ), human monocyte‐derived osteoclasts (Millan‐Linares et al, ), and THP‐1 cells (Millan‐Linares et al, ) at the same concentrations.…”

“…As depicted in Figure , the amino acidic sequence was described as glycine–proline–glutamate–threonine–alanine–phenylalanine–leucine–arginine (GPETAFLR) with α‐helix three‐dimensional structure (protein structure model was generated by the automated SWISS‐MODEL homology modeling pipeline, Bienert et al, ). Previously, antioxidative and anti‐inflammatory properties of GPETAFLR have been tested in a human myeloid lineage (Millan‐Linares et al, ). GPETAFLR showed decrease the expression of pro‐inflammatory cytokines as TNF‐α or IL‐1β and increase those that participate in the anti‐inflammatory response as IL‐10 or IL‐4 in THP‐1‐derived macrophages (Millan‐Linares et al, ), monocyte‐derived osteoclasts (Millan‐Linares et al, ), and primary human monocytes (Montserrat‐de la Paz et al, ).…”

“…Previously, antioxidative and anti‐inflammatory properties of GPETAFLR have been tested in a human myeloid lineage (Millan‐Linares et al, ). GPETAFLR showed decrease the expression of pro‐inflammatory cytokines as TNF‐α or IL‐1β and increase those that participate in the anti‐inflammatory response as IL‐10 or IL‐4 in THP‐1‐derived macrophages (Millan‐Linares et al, ), monocyte‐derived osteoclasts (Millan‐Linares et al, ), and primary human monocytes (Montserrat‐de la Paz et al, ).…”

A lupine (Lupinus angustifolious L.) peptide prevents non-alcoholic fatty liver disease in high-fat-diet-induced obese mice

“…Its immunomodulatory and anti-inflammatory capacities have been demonstrated. 11 Its effects have been previously investigated in macrophage THP-1-line derivatives, 11 osteoclasts derived from human primary monocytes, 12 and primary human monocytes. 13 Hence, it is an interesting compound to treat or prevent diseases characterized by inflammation.…”