INTRODUCTION: Clostridioides difficile, a leading cause of healthcare-associated infections, causes significant morbidity and mortality. Its pathogenesis centers on TcdA and TcdB toxins, which disrupt intestinal integrity, trigger inflammation, and promote extensive neutrophil infiltration. OBJECTIVE: The main objective of this study was to evaluate the role of PMNs in CDI using neutrophil depletion in a murine-ileal-ligated loop. METHODS: Mice were treated with C. difficile toxins TcdA, TcdB, and TcdBv, with PMN depletion achieved via intraperitoneal injections of Ly6G/Ly6C antibody. Histopathological analysis, cytokine quantification, and MPO activity assays were performed to assess the inflammatory and tissue damage responses. RESULTS: PMN depletion significantly reduced histopathological damage and proinflammatory responses. TcdA induced the highest inflammation and epithelial damage, while TcdB showed lower activity, except for MPO. TcdBv CONCLUSION: Our results show that PMN depletion reduced inflammatory responses and tissue damage, highlighting potential therapeutic strategies targeting PMN regulation. Further research on PMN extracellular traps (NETs) and their role in CDI is necessary to develop comprehensive treatments. Future studies should focus on combined in vivo and in vitro approaches to fully understand the pathological mechanisms and identify effective biomarkers for CDI therapy.