This study compared the protective effect of sodium selenite (SS) and selenomethionine (SeMet) on heat stress (HS) invoked porcine IPEC-J2 cellular damage and integrate potential roles of corresponding selenoprotein. Cells were cultured at 37 oC until 80% conﬂuence, and then subjected to four different conditions for 24 h: at 37 oC (Control), 41.5 oC (HS), 41.5 oC supplied with 0.42 µmol Se/L SS (SS) or SeMet (SeMet). HS significantly decreased cell viability, up-regulated mRNA and protein levels of HSP70, and down-regulated mRNA and protein levels of tight junction-related proteins (CLDN-1 and ZO-1). HS induced cells injury was associated with the up-regulation (P < 0.05) of 6 inflammation-related genes and 14 selenoprotein encoding genes, and down-regulation (P < 0.05) of 2 inflammation-related genes and 5 selenoprotein encoding genes. Compared with the HS group, SS and SeMet supplementation resulted in an increase (P < 0.05) in cell viability, decreases (P < 0.05) mRNA expression of HSP70 and 6 inflammation-related genes, and rescue (P < 0.05) of mRNA and protein levels of CLDN-1 and ZO-1. SS and SeMet supplementation changes expressions of 19 selenoprotein encoding genes in cells affected by HS. Both Se supplementation significantly recovered the protein level of GPX1 and increased SELP in the IPEC-J2 cells under HS, respectively. In summary, Se supplementation alleviated the negative impact of HS on IPEC-J2 cells, and their cellular protective effect was associates with regulation expression of selenoproteins, SeMet exhibited a better protective effect.