Effect of PI3K/AKT Pathway Modulation Mediated by Oncostatin M (OSM) on theProliferation, Migration, and Extracellular Matrix Gene Expression of Donkey SkinFibroblasts

Abstract

Donkeys exhibit a faster healing rate of dorsal skin wounds compared to their closely related counterparts, horses.This accelerated healing is accompanied by adequate collagen production without pathological over-deposition.However, studies exploring the underlying mechanisms of this superior wound healing in donkeys remain scarce.To explore the effects of OSM on the proliferation, migration, and extracellular matrix (ECM) gene expression of donkey skin fibroblasts, primary donkey skin fibroblasts were cultured and divided into four groups: OE-OSM, OE-NC, sh-OSM-2, and sh-NC.Cell viability was assessed using the Cell Counting Kit-8 (CCK-8) assay, and migration was evaluated through wound healing assays.Protein expression levels of collagen I (COL1), collagen III (COL3), and related factors were determined by Western blot (WB) analysis.Gene expression changes of COL1A1, COL3A1, Fibronectin (FN), and AKT Serine/threonine kinase 1 (AKT1) were quantified using RT-qPCR.Results indicated that OSM overexpression significantly reduced the proliferation and migration of donkey skin fibroblasts (P<0.01), while gene silencing produced opposite effects.Furthermore, overexpression of OSM notably reduced the expression levels of COL1A1, COL3A1, FN, and AKT1 (P<0.01), while simultaneously increasing Caspase-3 levels (P<0.01).These results suggest that OSM overexpression inhibits collagen synthesis (COL1 and COL3), reduces fibroblast proliferation and migration, and inhibits the PI3K/AKT signaling pathway.