Kaur Harmanpreet, Siraki Arno G, Uludağ Hasan, Dederich Douglas N, Flood Patrick, El-Bialy Tarek
Department of Dentistry, University of Alberta, Edmonton, Alberta Canada.
Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Alberta, Canada.
Ultrasound Med Biol. 2017 Nov;43(11):2699-2712. doi: 10.1016/j.ultrasmedbio.2017.07.002. Epub 2017 Aug 12.
We evaluated the activation of mitogen-activated protein kinase (MAPK) activation through reactive oxygen species (ROS) by application of low-intensity ultrasound (LIPUS) to MC-3 T3 E1 pre-osteoblasts. The cells were subjected to one LIPUS application for either 10 or 20 min, and the control group was exposed to a sham transducer. For ROS inhibition, 10 μM diphenylene iodonium (DPI) was added to the cells an hour before LIPUS application. Samples were collected 1, 3, 6, 12 and 24 h after LIPUS application, and cells were evaluated for ROS generation, cell viability, gene expression and MAPK activation by immunoblot analyses. LIPUS caused a significant increase in ROS and cell viability in the non-DPI-treated group. Expression of RUNX2, OCN and OPN mRNA was higher in the LIPUS-treated groups at 1 h in both the DPI-treated and non-DPI-treated groups; RUNX2 and OCN mRNA levels increased at 6 h. ERK1/2 activation was increased in the LIPUS-treated groups. These results indicate that LIPUS activates MAPK by ROS generation in MC-3 T3 E1 pre-osteoblasts.
我们通过对MC-3 T3 E1前成骨细胞施加低强度超声（LIPUS），评估了丝裂原活化蛋白激酶（MAPK）通过活性氧（ROS）的激活情况。将细胞进行一次LIPUS处理，处理时间为10或20分钟，对照组暴露于假换能器。为抑制ROS，在LIPUS处理前一小时向细胞中加入10μM二苯基碘鎓（DPI）。在LIPUS处理后1、3、6、12和24小时收集样本，并通过免疫印迹分析评估细胞的ROS生成、细胞活力、基因表达和MAPK激活情况。在未用DPI处理的组中，LIPUS导致ROS和细胞活力显著增加。在DPI处理组和未用DPI处理组中，LIPUS处理组在1小时时RUNX2、OCN和OPN mRNA的表达均较高；RUNX2和OCN mRNA水平在6小时时增加。LIPUS处理组中ERK1/2的激活增加。这些结果表明，LIPUS通过在MC-3 T3 E1前成骨细胞中产生ROS来激活MAPK。
