Xu Tianhua, Gu Jia, Li Chenghai, Guo Xiasheng, Tu Juan, Zhang Dong, Sun Wei, Kong Xiangqing
Department of Cardiology, The First Affiliated Hospital of Nanjing Medical UniversityNanjing 210029, Jiangsu, China.
Key Laboratory of Modern Acoustics, Department of Physics, Collaborative Innovation Center of Advanced Microstructure, Nanjing UniversityNanjing 210093, Jiangsu, China.
Am J Transl Res. 2018 Mar 15;10(3):948-956. eCollection 2018.
Low-intensity pulsed ultrasound (LIPUS) has been used widely in clinical therapy for bone fracture and soft tissue injury. However, whether LIPUS regulates primary preadipocyte function and adipogenesis remains unknown. In this study, we investigated the potential role of LIPUS in regulating visceral preadipocyte function. Resuspended rat visceral preadipocytes were treated with LIPUS (0.5 MHz, 109.44 mW/cm) for 1 min and then cultured for an additional 48 hours. Cell proliferation was examined using the CCK-8 assay, and the early apoptosis rate was determined by flow cytometry. In addition, we evaluated the related signaling pathway via examination of proliferating cell nuclear antigen (PCNA), peroxisome proliferator-activated receptor gamma (PPARγ), Bcl2, Bax, cleaved caspase 3 (C-C3), and mitogen-activated protein kinase (MAPK) member protein levels using western blot or quantitative real-time PCR (qRT-PCR). LIPUS inhibited preadipocyte proliferation and induced cell apoptosis. The protein expression of proliferation markers decreased, while expression of the apoptosis-related modulators increased following LIPUS treatment. LIPUS treatment decreased extracellular signal-regulated kinase (ERK) phosphorylation and increased p38 MAPK phosphorylation. Inhibition of p38 MAPK rescued the LIPUS-induced proliferation inhibition and apoptosis induction. Thus, treatment of rat visceral preadipocytes with 0.5 MHz LIPUS suppresses proliferation and promotes apoptosis via activation of p38 MAPK signaling.
低强度脉冲超声（LIPUS）已广泛应用于骨折和软组织损伤的临床治疗。然而，LIPUS是否调节原代前脂肪细胞功能和脂肪生成仍不清楚。在本研究中，我们调查了LIPUS在调节内脏前脂肪细胞功能中的潜在作用。将重悬的大鼠内脏前脂肪细胞用LIPUS（0.5MHz，109.44mW/cm）处理1分钟，然后再培养48小时。使用CCK-8法检测细胞增殖，通过流式细胞术测定早期凋亡率。此外，我们通过蛋白质印迹或定量实时PCR（qRT-PCR）检测增殖细胞核抗原（PCNA）、过氧化物酶体增殖物激活受体γ（PPARγ）、Bcl2、Bax、裂解的半胱天冬酶3（C-C3）和丝裂原活化蛋白激酶（MAPK）成员蛋白水平来评估相关信号通路。LIPUS抑制前脂肪细胞增殖并诱导细胞凋亡。LIPUS处理后，增殖标志物的蛋白质表达降低，而凋亡相关调节因子的表达增加。LIPUS处理降低细胞外信号调节激酶（ERK）磷酸化并增加p38 MAPK磷酸化。抑制p38 MAPK可挽救LIPUS诱导的增殖抑制和凋亡诱导。因此，用0.5MHz LIPUS处理大鼠内脏前脂肪细胞可通过激活p38 MAPK信号通路抑制增殖并促进凋亡。
