Abstract Di(2-ethylhexyl)phthalate (DEHP), is a toxic environmental pollutant in our life which can contaminate air, water, and soil. The hepatoprotective effect of silymarin on DEHP-induced injury in FL83B mouse liver cells was investigated by analyzing the cell viability, lactate dehydrogenase (LDH), alanine aminotransferase (ALT), cell cycle arrest, and cell morphology. The results revealed that cell viability decreased while released LDH and ALT increased with the increase of DEHP concentrations. Moreover, cell population of sub-G1 and S phase increased as the concentrations of DEHP increased. Silymarin at 25μM achieved the highest hepatoprotective effect and exhibited 79% cell viability while only 46% cell viability was found in DEHP injured control. It was also found to reduce LDH release and cell populations of sub-G1 and S phase. Therefore, silymarin could ameliorate DEHP-induced injury and have potential to be further developed as a natural ingredient of health food against phthalate plasticizers induced liver injury.