The present study aimed to investigate the in uence of heat stress on prostaglandin production and the expression of key genes, including COX-2, PGES, PGFS, ITGAV, and LGALS15, in buffalo endometrial epithelial cells. Buffalo genitalia, containing ovaries with corpus luteum (CL), were collected immediately post-slaughter and transported to the laboratory. The stages of estrous cycle were determined based on macroscopic observations of the ovaries, and uteri from the mid luteal phase (days 6-10 of the estrous cycle) were utilized. Uterine lumens were washed and treated with trypsin to isolate epithelial cells, which were then cultured at control temperature (38.5°C for 24 hr) or exposed to elevated temperatures (38.5°C for 6 hr, 40.5°C for 18 hr; HS). The supernatant and endometrial epithelial cells were collected at various time points (0, 3, 6, 12, and 24 hr) from both control and treatment group. Despite the elevated temperature (40.5°C) not affecting the in vitro production of PGF 2α and PGE 2 in epithelial cells, there was a signi cant (P < 0.05) increase in COX-2, PGES, and PGFS transcripts in response to heat stress.Interestingly, the expression of ITGAV and LGALS15 mRNAs in endometrial epithelial cells remained unaltered under elevated temperature conditions. In conclusion, this study suggests that while elevated temperature did not directly modulate prostaglandin production, it promoted the expression of COX-2, PGES, and PGFS mRNA in buffalo endometrial epithelial cells. These ndings contribute to our understanding of the molecular responses of buffalo reproductive cells to heat stress, potentially informing strategies for mitigating the impact of environmental factors on reproductive outcomes in this species.