Abstract
Skin wound healing is an important fundamental problem in biological and medical fields. This study aimed to investigate wound healing promotion of protein extract from tail-amputated Eisenia foetida (E. foetida) and reveal the mechanism correlated with the Ras/Raf/MEK/ERK signaling pathway. Proteins extracted from tail-amputated E. foetida were applied on rats' full-thickness excisional wounds to evaluate their regenerative efficacy. Rat skin tissues around surgical defects were analyzed by immunofluorescence staining and Western blot methods. The Ras/Raf/MEK/ERK signaling pathway was further investigated in vitro using the NIH3T3 cell line. A tail-amputated protein extract (ES2) from E. foetida significantly accelerated rat wound healing ability via higher re-epithelialization and ECM deposition in the tissue section compared to the blank control and un-amputated earthworm extract groups. Furthermore, ES2 treatment dramatically accumulated the expressions of platelet-derived growth factor (PDGF), transforming growth factor-β (TGF-β), and hydroxyproline (HYP) in wound areas on day 7 without their accumulation on day 21 post-wounding, diminishing excessive scar formation. Accelerated wound healing ability with the ES2 was proved to correlate with the up-regulation of the Ras/Raf/MEK/ERK signaling pathway. The mRNA expression of this pathway increased significantly in NIH3T3 cells after being treated with the ES2 at an appropriate concentration. The tail-amputated E. foetida proteins (ES2) can significantly promote skin wound healing better than the un-amputated earthworm tissue extract without excessive scar tissue formation. This effect was related to the up-regulation of the Ras/Raf/MEK/ERK signaling pathway.