Pea protein-derived tripeptide LRW shows osteoblastic activity on MC3T3-E1 cells via the activation of Akt/Runx2 pathway

Osteoporosis is a bone disease affecting 2 million people comprising of 1 in 3 women and 1 in 5 men in Canada. One possible approach to prevent this disease is to stimulate the activity of osteoblasts (bone forming cells) using food protein-derived bioactive peptides. In previous study, we identified a tripeptide LRW (Leu-Arg-Trp) from pea protein, which exerts ACE inhibitory activity. The aim of this work is to investigate the effect of LRW on promoting osteoblastic activity. The tripeptide LRW treatment (50 µM) in MC3T3-E1 cells increased cell proliferation (4-fold increase) by BrdU incorporation assay. Moreover, we found that tripeptide LRW stimulated osteoblastic differentiation by increased levels of type 1 collagen (COL1A2; 3-fold increase), alkaline phosphatase (ALP; 4-fold increase), runt-related transcription factor 2 (RUNX2; 2-fold increase). Furthermore, tripeptide LRW increased matrix mineralization as evidenced by Alizarin-S red staining and nodule formation, osteoprotegrin levels (OPG; 2-fold increase) and thus decreasing bone resorption, and the wound healing based on cell migration assay. The pea protein derived bioactive peptide LRW can positively modulate the activity of osteoblasts, indicating its potential use for the prevention of osteoporosis.