Abstract: Objective：To discuss the effect of 1,25(OH)2D3 in regulation of the level of oxidative stress in temporomandibular joint chondrocyte by constructing the model of 1,25(OH)2D3 deficiency chondrocyte in oxidative stress state. Methods：Knocking down the level of vitamin D receptor in the temporomandibular joint condylar chondrocytes of SD rats by lentivirus infection in order to simulate the 1,25(OH)2D3 deficiency status. Using the exogenous hydrogen peroxide to stimulate both the normal and 1,25(OH)2D3 deficiency chondrocytes we established the oxidative stress model in vitro. Detecting the alteration of oxidative stress level in both groups after being treated with exogenous 1,25(OH)2D3, Malonaldehyde, total superoxide dismutase, Glutathione peroxidase and reactive species oxygen in each group were detected and compared as representing the oxidative stress level in each group. Results：The oxidation level was increased in 1,25(OH)2D3 deficiency chondrocytes after, being treated with hydrogen peroxide. Treated with 1,25(OH)2D3, the level of oxidative stress were decreased obviously in both the normal and 1,25(OH)2D3 deficiency chondrocytes. However the level of oxidative stress in 1,25(OH)2D3 deficiency chondrocytes was still fairly higher than that of the normal group. Conclusions：The deficiency of 1,25(OH)2D3 can increase the level of oxidative stress.