A non-bactericidal glycine-rich peptide enhances cutaneous wound healing in mice via the activation of the TLR4/MAPK/NF-κB pathway

Although the antibacterial properties of glycine-rich peptides from prokaryotes to eukaryotes have been well characterized, their role in skin wound healing remains poorly understood, especially non-bactericidal glycine-rich peptides. Herein, a novel glycine-rich (46.5%) peptide (Smaragin, SRGSRGGRGGRGGGGRGGRGRSGSGSSIAGGGSRGSRGGSQYA) was identified from the skin of the tree frog Zhangixalus smaragdinus. Unlike other glycine-rich peptides, Smaragin showed no antimicrobial activity in vitro but significantly enhance wound healing in full-thickness dermal wounds in mice. In comparison with other wound healing-promoting peptides, Smaragin did not directly affect the proliferation and migration of keratinocytes, vascular endothelial cells, and fibroblasts. However, it notably increased phagocytes infiltration at the wound site by 0.5-day post-injury. Smaragin was not a direct chemoattractant for phagocytes, but it stimulated macrophages to secrete chemokines CXCL1 and CXCL2, which indirectly enhanced the migration of phagocytes, keratinocytes and vascular endothelial cells. Moreover, Smaragin promoted the polarization of macrophages from a pro-inflammatory M1-type to an anti-inflammatory M2 phenotype at the wound, which is associated with angiogenic activity. As expected, CD31, the most common analyzed marker of angiogenesis, showed a significant increase in vascular network area. Subsequent studies revealed that Smaragin promoted the chemokine level and polarization of macrophages via the TLR4/MAPK/NF-κB pathway, which enhanced the number of phagocytes and the regeneration of the epidermis and blood vessels at the wound, thereby accelerating skin wound healing in mice. These findings highlight the skin healing properties of non-bactericidal glycine-rich peptides and display the potential of Smaragin as a promising candidate for developing effective wound healing therapies.