Objective：This study aims to investigate if low intensity pulsed ultrasound（LIPUS）has the inhibition effect of on calcification of rat vascular smooth muscle cells（rVSMC）and rat vascular adventitial fibroblasts（rVAF）induced by hypercalcemia and hyperphosphatemia. Methods：RVSMCs and rVAF were extracted，calcium quantification，alkaline phosphatase （ALP） activation and Alizarin red staining to detect the different intensity of LIPUS on calcification；Western blot and real-time PCR were performd to explore the effect of LIPUS on expression of osteogenic markers. Results：LIPUS treatment significantly inhibited the calcium quantification，ALP activation of rVSMCs and rVAF induced by hypercalcemia and hyperphosphatemia. Meanwhile，the expression of osteogenic markers of rVSMCs and rVAF decreased after 14 mW/cm2 LIPUS treatment. Conclusion：LIPUS treatment could inhibit the expression of osteogenic markers in both rVSMCs and rVAF induced by hypercalcemia and hyperphosphatemia. Therefore，LIPUS has protection effect on rVSMCs and rVAF calcification caused by hypercalcemia and hyperphosphatemia.