Background: Intestinal epithelial injury stress is one of the most common stresses in the contemporary poultry breeding. This study aims to evaluate the protective effects of Methylsulfonylmethane (MSM) and epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI, erlotinib) on injury stress induced by lipopolysaccharides (LPS) in chicken intestinal epithelial cells (IECs). Methods: Cell Counting Kit-8 (CCK-8) and EdU assays were used to evaluate cell viability and proliferation. The cell cycle was analyzed by flow cytometry method. The mRNA transcript and protein expression levels of EGFR were detected by RT-qPCR and Western blotting (WB). The levels of pro-inflammatory cytokines were determined by using ELISA kits. Result: The results have shown that MSM could promote the cell proliferation and regulate the cell cycle arrest when EGFR inhibited (P less than 0.05). In addition, both MSM and EGFR-TKI could decrease the expressions of IL-1β, IL-6 and TNFα (P less than 0.05). These results showed that the beneficial roles of MSM in chicken IECs induced by LPS may be related to promoting cell proliferation and inhibiting inflammation; meanwhile, our experimental results may provide valuable information for further study of the possible mechanism of MSM to improve the chicken intestinal epithelial function by regulating EGFR signaling pathway.