Objective To establish a palmitic acid-induced bone formation inhibition model of zebrafish lipotoxic injury. Methods The embryos of AB strain zebrafish were randomly divided into blank Control group (Control group), palmitic acid group (PA group) and simvastatin group (SIM group). From 3 dpf (days post fertilization), PA and SIM groups were given PA . From 5 dpf, the SIM group was administered continuously for 4 days. Calcein staining, Nile red staining, triglyceride and total cholesterol content determination and q-PCR to determine whether the model was successfully established on the 9 dpf. Results PA significantly decreased the number of vertebrae, promoted lipid accumulation, increased triglyceride triglyceride and total cholesterol content, promoted lipid-related genes and inhibited the expression of osteogenic genes. SIM can improve the inhibitory effect of PA on bone formation in zebrafish. Conclusion PA can successfully create the inhibition model of lipotoxic injury bone formation similar to the pathological process of OP, and the method is simple, sensitive and controllable, which can be used for drug screening of OP and related diseases.