Objective: The aim of this study was to investigate the potential effects and mechanisms of low-intensity pulsed ultrasound (LIPUS) on the metabolism of hypertrophic adipose tissue induced by high-fat diet. Methods: Twenty male mice fed a 60% kcal high-fat diet (HFD group) for 10 weeks as an obesity model, while ten fed a 10% kcal low-fat diet (NC group) as a negative control. Then selected 10 mice randomly matched body weight with HFD group for LIPUS treatment at inguinal fat depot as a LIPUS group. After 10 weeks LIPUS irradiation, the whole blood, skin, muscles and inguinal fat pads at LIPUS region, liver, spleen and representative visceral fat depots. We evaluated the effects of LIPUS on the prevention of a series of disorders through an analysis of plasma biochemical profile, body and fat pads mass, adipocytes size, triglyceride contents and associated genes and proteins expression in inguinal adipose depot. The mRNA expression levels of transcription factors for adipogenic differentiation (PPAR-γ and C/EBPα), transcription factors for ATGL (IRF4, FoxO1), lipolyases (ATGL and HSL) and adipokines (ADPN and LEP) genes in the target region were analyzed by qPCR. Additionally, Western blot was applied to detect the protein levels of C/EBPα, ATGL, HSL and related signaling pathways (PKA and ERK1/2). Furthermore, safety tests on LIPUS were conducted on the basis of complete blood counts and tissue sections at LIPUS site stained with hematoxylin-eosin. Results: LIPUS reduced local adipose hypertrophy at irradiation regions in obese mice through downregulating adipogenic differentiation related transcription factors and upregulating lipolytic enzymes expression and associated transcription factors. In addition, LIPUS stimulated lipolysis via activating PKA and ERK1/2 to phosphorylate hormone sensitive lipase. Conclusion: LIPUS diminishes local adipose tissue hypertrophy induced by high-fat diet through inhibiting adipogenic differentiation and encouraging lipolysis procedure.