Effects of Low‐Intensity Pulsed Ultrasound on the Regulation of Free Fatty Acid Release in 3T3‐L1 Cells

Objectives To investigate the effects of low‐intensity pulsed ultrasound (LIPUS) on the proliferation, differentiation, and tumor necrosis factor‐α (TNF‐α)‐induced lipolysis of 3T3‐L1 cells, and to explore the feasibility of regulating the release of free fatty acids (FFA) to prevent lipotoxicity. Methods Different intensities (30, 60, 90, and 120 mW/cm 2 ) of LIPUS were applied to 3T3‐L1 preadipocytes for different durations (5, 10, 15, 20, 25, and 30 minutes). Appropriate parameters for subsequent experiments were selected by assessing cell viability. The effect of LIPUS on the proliferation and differentiation of 3T3‐L1 cells was evaluated by microscope observation, flow cytometry, and lipid content determination. After treated with LIPUS and TNF‐α (50 ng/mL), the degree of lipolysis was assessed by measuring the extracellular FFA content. Quantitative real‐time polymerase chain reaction (qRT‐PCR) was used to detect the mRNA expression of relevant genes. Results Different parameters of LIPUS significantly enhance the viability of 3T3‐L1 cells ( P < .05), with 20 minutes and 30 mW/cm 2 as the most suitable settings. After LIPUS treatment, 3T3‐L1 cell proliferation accelerated, apoptosis rate and G1 phase cell proportion decreased, the content of lipid droplets and TG was increased in differentiated cells, while FFA release decreased ( P < .05). The expression of PCNA, PPARγ, C/EBPα, Perilipin A mRNA increased, and the expression of TNF‐α, ATGL, HSL mRNA decreased ( P < .05). Conclusions LIPUS could promote the proliferation and differentiation of 3T3‐L1 cells and inhibit TNF‐α‐induced lipolysis, indicating its potential as a therapy for mitigating lipotoxicity caused by decompensated adipocytes.