Objective To investigate the effect of hypoxia inducer cobalt dichloride (CoCl₂ ) on autophagy of 3T3-L1 adipocytes. Methods 3T3-L1 adipocytes were cultured and induced to mature adipocytes. CoCl₂ was used as an inducer of hypoxia in vitro. Mature adipocytes were divided into control and CoCl₂ treatment groups with various concentrations and times. In accordance with the result, 150 μmol/ L cobalt dichloride was selected to treat adipocytes for 0, 12, 24 and 48 h. Then, the cells were collected for related tests. MTT assays were used to assess cell survival. Expression of HIF-1α, autophagy-related protein LC3-Ⅱ, Beclin-1 and Glut-1 was analyzed by Western blot. The level of autophagy was measured by immunofluorescence. TNF-α and IL-6 levels in culture supernatants of adipocytes were measured by ELISA. Results 150 μmol/ L CoCl₂ is the optimal intervention concentration to regulate the autophagy level of 3T3-L1 adipocytes. The levels of autophagic activity increased without reduction in cell viability after 150 μmol/ L CoCl₂ intervention in mature adipocytes for 24 h. The expression protein levels of HIF-1α, LC3-Ⅱ, Beclin-1 and Glut-1 were significantly increased in adipocytes. However, the secretion level of inflammatory factors TNF-α and IL-6 did not increased significantly. The level of autophagic activity and cell viability were decreased at 48 h. The expression protein levels of HIF-1α, LC3-Ⅱ, Beclin-1 and Glut-1 were significantly decreased in adipocytes. However, the increased secretion of inflammatory cytokines TNF-α and IL-6 were observed. Conclusions Autophagy was moderately activated in adipocytes by 150 μmol/ L CoCl₂ treatment for 24 hours. Autophagy was activated in a HIF-1α-dependent manner, which plays a role in protecting adipocytes from inflammatory damage and improving insulin resistance.