Collagen from Iris squid grafted with polyethylene glycol and collagen peptides promote the proliferation of fibroblast through PI3K/AKT and Ras/RAF/MAPK signaling pathways

Abstract

Collagens from marine sources have been used widely in food, cosmetics and tissue engineering application due to their excellent functional and biological properties. In the present study, a novel protein, collagen from iris squid skin (SSC) was characterized, grafted with polyethylene-glycol (PEG) and Acid-Green 20 (AG) and was investigated the molecular signaling pathways in L-929 fibroblast cells along with their structural peptide analogs. SDS-PAGE and IR spectrum of SSC analysis showed the typical structure of type I collagen. The fibroblast proliferation was evaluated for SSC, SSC grafted PEG (SSC-PEG) and their structural analogs including Gly-Pro-Leu-Gly-Leu-Leu (PEP1), Gly-Pro-Leu-Gly-Leu-Leu-Gly-Phe-Leu (PEP2), Gly-Pro-Leu-Gly-Leu-Leu-Gly-Phe-Leu-Gly-Pro-Leu (PEP3) and Gly-Pro-Leu-Gly-Leu-Leu-Gly-Phe-Leu-Gly-Pro-Leu-Gly-Leu-Ser (PEP4). The optimal concentration of SSC and its derivative was 0.07 μ mol/L. The fibroblast growth-promoting factors were promoted by all the treatment groups by accelerating the PI3K/AKT and Ras/RAF/MAPK signaling pathways in L-929 cells, and inhibiting the secretion of apoptotic factors. Compared to the control group, mRNA and protein expression of AKT in the PI3K/AKT and Ras in Ras/RAF/MAPK signaling pathway were accelerated significantly by PEP4, respectively, while the Bax value was significantly lower (P < 0.01). The promoting effect of PEP1, PEP2, PEP3 and PEP4 on L-929 cells was closely related to the length of the peptides. Therefore, this study disclosed that PEP1, PEP2, PEP3 and PEP4 were novel analogs that greatly promote the proliferation of L-929 cells through PI3K/AKT and Ras/RAF/MAPK signaling pathways.