This study aims to investigate the impact of curcumin (CU) supplementation in semen extender on post/thawed rabbit sperm quality, and how curcumin prevents spermatozoa from harm during the preservation process. Semen was diluted with (TCG) extender supplemented with curcumin (CU) at levels of (CU 0, CU 0.25, CU 0.5 , and CU 1 mmol, respectively) compared to (TCG) extender supplemented with 200 µm of Trolox as a positive control, and the following parameters were analyzed: sperm motility, membrane integrity, acrosomal intact, viability, biochemical evaluation of antioxidants indices, mitochondrial activity, (ROS) level, DNA fragmentation, and expression level of selected antioxidant-related genes  and antifreeze related gene using RT-qPCR. Our results revealed that groups of (CU 0.5 mmol, CU 0.25 mmol and 200 µm Trolox) have a positive influence on post-thawed sperm progressive motility with no significant difference. Moreover, CU 0.5 mmol group significantly (p ≤ 0.05) improved antioxidants enzymes values compared to all groups while malondialdehyde MDA reduced in (CU 0.5 mmol, CU 0.25 mmol and 200 µm Trolox groups) in comparison with (control and CU 1mmol) groups. The CU 0.5 mmol group exhibited significantly (p ≤ 0.05) higher mitochondrial membrane potential activity, lower ROS accumulation and DNA fragmentation levels than other groups, Antioxidant-related genes and HSP90 were significantly (p ≤ 0.05) up-regulated in sperm cryopreserved in CU 0.5 mmol group compared to other groups. This study concluded that supplementation of curcumin with concentration (0.5 mmol) promotes Sperm quality characteristics, reduces ROS, up regulates antioxidants and antifreeze related genes.