Introduction: Females seem be more tolerant to liver ischemia/reperfusion (I/R) injury than males, the mechanisms of which remains unknown. The objective of the current study was to identify the role of sex-determining region on Y chromosome (SRY) on hepatic I/R-induced liver damage. Method: 771 patients who underwent hepatic resection (HR) for hepatic tumors were included, and the postoperative outcome was compared between female and male patients by using propensity score matching (PSM) analysis. Hepatocyte-specific SRY transgenic mice were generated and subjected to an I/R model in vivo. In vitro experiments were performed by treating primary hepatocytes from these mice with hypoxia/reoxygenation stimulation. The function of SRY in I/R-induced liver damage and the potential underlying mechanisms were investigated through various phenotypic analyses and biological approaches. Result: The incidence of overall postoperative complications, mortality and liver failure was significantly higher among male versus female patients (all p<0.05). SRY expression was significantly upregulated after hepatic I/R injury. Overexpression of SRY in male or female (SRY TG) mice promoted hepatic I/R injury as manifested by increased inflammatory reaction, oxidative stress and cell death versus age- and gender matched littermates. Direct binding and activation of GSK3β by SRY and subsequent inhibition of Wnt/β-catenin signaling accounted for the susceptibility of SRY TG mice to hepatic I/R injury. Activation of β-catenin by LiCl abrogated SRY-initiated I/R injury. Conclusion: SRY is a novel mediator inducing sex specific hepatic I/R via inactivation of Wnt/β-catenin signaling. Modulation of SRY in males is a unique mode of regulating Wnt/ β-catenin signaling, and provides novel opportunities in devising therapeutics in specific hepatic injury.