The effects of photobiomodulation therapy on mouse pre-osteoblast cell line MC3T3-E1 proliferation and apoptosis via miR-503/Wnt3a pathway
The effects of photobiomodulation therapy on mouse pre-osteoblast cell line MC3T3-E1 proliferation and apoptosis via miR-503/Wnt3a pathway
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DOI：
10.1007/s10103-018-2636-0
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发表时间：
2019-04-01
影响因子：
2.1
通讯作者：
Li, Meihua
中科院分区：
文献类型：
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作者：
Li, Qiushi;Li, Chen;Li, Meihua
光生物调节疗法（PBMT）已被证明是调节成骨细胞增殖。microRNAs（miRNAs）参与成骨细胞的多种病理生理过程，但miRNAs在PBMT促进成骨细胞增殖中的作用尚不清楚。本研究旨在探讨PBMT治疗的效果（3.75J/cm（2））通过miR-503/Wnt 3a通路对小鼠前成骨细胞系MC 3 T3-E1增殖和凋亡的影响;同时检测PBMT处理后miR-503和Wnt 3a的表达，以及miR-503对Wnt信号分子Wnt 3a、β-catenin、Runx 2、凋亡蛋白caspase-3 Bcl-2的表达。PBMT参数为：连续波长808 nm，输出功率0.401W，功率密度0.042W/cm（2），光斑尺寸9.6cm（2），能量36 J，能量密度3.75J/cm（2），照射时间90 s，每12 h照射3次，光源距离14.5cm，激光束发散角7 °。本研究通过生物信息学分析和荧光素酶报告基因分析，预测并验证了靶点关系。通过qPCR和western blot分析检测基因mRNA和蛋白质表达。MTT法检测miR-503对MC 3 T3-E1细胞增殖的影响。流式细胞术检测细胞凋亡。结果显示，PBMT处理降低了miR-503的表达，提高了Wnt 3a的水平（p
Photobiomodulation therapy (PBMT) has been demonstrated as regulating osteoblast proliferation. MicroRNAs (miRNAs) are involved in various pathophysiologic processes in osteoblast, but the role of miRNAs in the PBMT-based promotion of osteoblast proliferation remains unclear. This study aimed to investigate the effects of PBMT treatment (3.75J/cm(2)) on mouse pre-osteoblast cell line MC3T3-E1 proliferation and apoptosis via the miR-503/Wnt3a pathway; meanwhile, detect the expressions of miR-503 and Wnt3a after PBMT treatment and the role of miR-503 in regulating Wnt signaling molecules Wnt3a, -catenin, Runx2, apoptotic proteins caspase-3, and Bcl-2 in vitro. The PBMT parameters were as follows: 808nm continuous wavelength, 0.401W output power, 0.042W/cm(2) power density, 9.6cm(2) spot size, 36J energy, 3.75J/cm(2) energy density, 90s irradiation for three times per 12h, 14.5cm distance of the laser source and the angle of divergence of the laser beam was 7 degrees. In our present study, the target relationship was predicted and verified by bioinformatics analysis and luciferase reporter assays. Gene mRNA and protein expressions were examined by qPCR and western blot analysis. The MTT method was used to evaluate the effect of miR-503 on MC3T3-E1 cells proliferation. And cell apoptosis was examined by flow cytometry. The results showed that PBMT treatment reduced the expression of miR-503 and increased the level of Wnt3a (p