Aim To observe the correlative mechanism between ghrelin and osteoprotegerin (OPG)/receptor activator of nuclear factor kappa B ligand (RANKL) in A7r5 calcification. Methods Changes of ghrelin in A7r5 calcification under high-lipid, glucose-coexisting conditions were observed. And then, effects of OPG and RANKL regulated by exogenous ghrelin on A7r5 calcification under high-lipid, glucose, β-glycerophosphate-coexisting conditions were investigated. Finally, the interaction between OPG/RANKL and ghrelin in A7r5 calcification under high-lipid, glucose, β-glycerophosphate-coexisting conditions was explored. Von Kossa staining, alizarin red staining, extracellular calcium deposition assay, immunohistochemical staining, Western blot analysis were performed after incubation of 14 days. ResultsVon Kossa staining, alizarin red staining and extracellular calcium deposition test showed that calcium deposition of A7r5 cells significantly increased by 5.21 times under diabetic metabolic disorder simulated by high-lipid, glucose, β-glycerophosphate-coexisting conditions. Ghrelin immunohistochemical staining and IPP6 quantitative analysis indicated that the expression of ghrelin was dramatically reduced with the formation and development of calcification. Exogenous ghrelin could increase the relative expression of OPG by 6.25 times. Compared with the high glucose,high lipid and calcified group, the calcium deposition in anti-OPG group and RANKL group were raised by 1.33 times (P<0.05) and 1.59 times (P<0.05), respectively. Compared with anti-OPG group, the calcium deposition in anti-OPG plus ghrelin group was decreased by 41.9% (P<0.05); Compared with RANKL group, the calcium deposition in RANKL plus ghrelin group was decreased by 57.8% (P<0.05). Conclusion Ghrelin regulated A7r5 cells calcification by OPG/RANKL signal.