Objective To establish a palmitic acid (PA)-induced model of lipotoxic bone formation inhibition in zebrafish. Methods AB strain zebrafish embryos were divided randomly into a blank control group, PA group, and simvastatin (SIM) group. Embryos in the PA and SIM groups received PA from 3 days post-fertilization ( dpf), and embryos in the SIM group received SIM continuously for 4 days from 5 dpf. Establishment of the model was confirmed at 9 dpf by calcein staining, Nile red staining, triglyceride and total cholesterol content determination, and q-PCR. Results PA significantly decreased the number of vertebrae, promoted lipid accumulation, increased triglyceride and total cholesterol contents, promoted the expression of lipid-related genes PPARγ、 c / EBPα, and inhibited the expression of osteogenic genes ALP and RUNX2. SIM improved the inhibitory effect of PA on bone formation in zebrafish. Conclusions PA can successfully create a lipotoxic model of bone-formation inhibition, similar to the pathological process of osteoporosis, using a simple, sensitive, and controllable method. This model can then be used for drug screening for osteoporosis and related diseases.