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2022, Vol. 26 ›› Issue (13): 2006-2011Low-intensity pulsed ultrasound promotes the proliferation of bone marrow mesenchymal stem cells by inducing cyclin D1 up-regulationLi Zhi1, 2, 3, Hua Yongyong4, Zhang Jianquan4, Fu Zhen4
Abstract: BACKGROUND: Previous studies found that low-intensity pulsed ultrasound can promote the proliferation of human bone marrow mesenchymal stem cells. OBJECTIVE: To investigate the mechanism of low-intensity pulsed ultrasound on the proliferation of human bone marrow mesenchymal stem cells via the FAK/PI3K/Akt signaling pathway. METHODS: The purchased commercial human bone marrow mesenchymal stem cells were resuscitated and cultured to the fourth generation, and they were divided into 0 mW/cm2 ultrasound group (control group), 50 mW/cm2 ultrasound group, and 50 mW/cm2 ultrasound + PF-562271 group. The 50 mW/cm2 ultrasound + PF-562271 group was pretreated with 1 μL FAK/PI3K/Akt signaling pathway inhibitor PF-562271 before ultrasound treatment, once every 24 hours, 5 minutes each time. After three stimuli, cell proliferation was detected by Cell Counting Kit-8 and the cell cycle was detected by flow cytometry. Western blot assay was used to detect the expression of FAK and its downstream target gene and cyclin D1. RESULTS AND CONCLUSION: (1) Compared with the control group, the cell proliferation ability was significantly enhanced in the 50 mW/cm2 ultrasound group (P < 0.05). After inhibiting the FAK/PI3K/Akt signaling pathway using PF-562271, the cell proliferation ability was significantly reduced (P < 0.001). (2) Compared with the control group, the expression levels of FAK, p-FAK, PI3K, p-PI3K, Akt, p-Akt and cyclin D1 were all up-regulated in the 50 mW/cm2 ultrasound group (P < 0.05). Compared with the 50 mW/cm2 ultrasound group, the above indicators were significantly reduced in the 50 mW/cm2 ultrasound + PF-562271 group (P < 0.05). (3) Compared with the control group, the proportion of cells in the G1 phase decreased in the 50 mW/cm2 ultrasound group (P=0.026), and the proportion of cells in the S phase increased (P=0.002). Compared with the 50 mW/cm2 ultrasound group, the proportion of cells in G1 phase increased (P=0.023), and the proportion of cells in S phase decreased in the 50 mW/cm2 ultrasound + PF-562271 group (P=0.035). (4) These results indicate that low-intensity pulsed ultrasound promotes the proliferation of human bone marrow mesenchymal stem cells via FAK/PI3K/Akt signaling pathway-induced cyclin D1 up-regulation. Key words: stem cells, human bone marrow mesenchymal stem cells, low-intensity pulsed ultrasound, proliferation, pathway, cell cycle |