This study aimed to investigate the tolerance and physiological differences between male and female turbot (Scophthalmus maximus) in response to hypoxic stress. Female (169.95±13.55) g and male (170.08± 19.02) g turbot of similar sizes were used to determine the concentration of dissolved oxygen at their critical oxygen tension (P_crit). Furthermore, changes in blood physiological and biochemical indexes, hepatic antioxidant activity, gas exchange rate, respiratory rate, and gill histomorphology were analyzed under hypoxic stress and again after reoxygenation. The results showed that the dissolved oxygen concentrations of male and female turbot under P_crit were (3.34±0.23) mg/L and (3.22±0.17) mg/L, respectively, with no significant difference observed (P＞0.05). Turbot plasma cortisol (COR) glucose (GLU) content, white blood cell (WBC) number, hepatic superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activity, and malondialdehyde (MDA) content were all significantly increased (P＜0.05) after 6 h of hypoxic stress treatment. Meanwhile, significant differences in CAT activity, GSH-Px activity, and MDA content were observed between males and females after treatment with hypoxia for 6 h. The gas exchange rate, respiratory rate, secondary lamellar length (SLL), interlamellar distance (ID), and perimeter of gill lamella of both male and female turbot were significantly increased, whereas the secondary lamellar width (SLW) of gill lamella were significantly decreased (P＜0.05). The clubbing and matrix hyperplasia at the end of gill lamella of female and male turbot manifested similar results to those of the SLL (P＜0.05), with no significant difference observed between the sexes. All aforementioned parameters were recovered to normal levels after reoxygenation for 12 h. These results demonstrate that there were physiological differences observed between male and female turbot under hypoxic stress conditions, especially in CAT activity, GSH-Px activity, and MDA content. These changes aimed to meet the metabolic demand of the organism while enhancing the activity of liver antioxidant enzymes to alleviate the oxidative stress damage caused by hypoxic stress. Notably, males exhibited greater tolerance to low oxygen levels than females.