Background: The relationship between platelet count and sepsis outcomes in intensive care units (ICUs) requires comprehensive investigation through large-scale multicenter studies. Methods: In this multicenter retrospective cohort study, we analyzed 17,977 sepsis patients from 208 U.S. hospitals (2014-2015) using the eICU Collaborative Research Database v2.0. Analyses were adjusted for demographics, clinical parameters, comorbidities, and treatments. Generalized additive models and two-piecewise linear regression were used to assess the relationship between platelet count and mortality. Results: A U-shaped relationship was identified with an inflection point at 176 x 109/L. Below this threshold, each 10 x 109/L increase in platelet count was associated with a 6% decrease in mortality risk (adjusted OR 0.94, 95% CI 0.93-0.95, p < 0.0001), while above it, each 10 x 109/L increase was associated with a 1% increase in mortality risk (adjusted OR 1.01, 95% CI 1.00-1.01, p = 0.0153). Conclusion: This large-scale, multicenter retrospective study has made a significant contribution to understanding the association between platelet count and mortality in patients with sepsis in intensive care units. We identified a critical threshold of 176 x 10(9)/L for platelet count and demonstrated a distinct U-shaped relationship with 30-day in-hospital mortality, providing valuable reference criteria for clinical risk stratification.