Tian Tian, Wang Ziting, Chen Leyi, Xu Wenan, Wu Buling
Department of Endodontics, Shenzhen Stomatology Hospital (Pingshan), Southern Medical University, 143 Dongzong Road, Pingshan District, Shenzhen, 518118, China.
Department of pediatric dentistry, Shenzhen Stomatology Hospital (Pingshan), Southern Medical University, 143 Dongzong Road, Pingshan District, Shenzhen, 518118, China.
Lasers Med Sci. 2023 Mar 18;38(1):86. doi: 10.1007/s10103-023-03753-x.
Macrophages are the main mediators of the inflammatory response and play a major role in the onset and maintenance of periodontitis. Studies revealed that photobiomodulation (PBM) can change the polarization state of macrophages and inflammation reduction, although the cellular mechanisms are not fully elucidated. Here, the present study explored the effect of PBM (980 nm) on undifferentiated and M1-type macrophages and the underlying mechanism. RAW264.7 cells were exposed to laser irradiation under different laser parameters (0.5, 5.0, and 10.0 J/cm) with or without LY294002 (an inhibitor of PI3K pathway). Then, confocal laser microscopy was used to observe cell differentiation; qPCR was performed to examine the gene expression and western blotting was used to detect the protein in the PI3K/AKT/mTOR pathway and activated macrophage markers. The obtained results revealed that 980 nm PBM increased the mRNA expression of iNOS, Il-10, Arg1, and Il-12 along with the inflammatory cytokines Tnfα, IL-1β, and Il-6 in M0-type macrophages in dose-dependent manner. More interestingly, PBM at 5 J/cm decreased the mRNA expression of iNOS, Il-12, Tnfα, IL-1β, and Il-6 and increased the expression of Arg1 and Il-10 by M1-type macrophages, along with the elevated expression of phosphorylation of AKT and mTOR. Moreover, PBM-induced M1-type macrophage polarization was significantly attenuated via LY294002 treatment. These suggest that 980 nm PBM could activate M0-type macrophages and increase M2/M1 ratio via the PI3K/AKT/mTOR pathway.
巨噬细胞是炎症反应的主要介质，在牙周炎的发生和维持中起主要作用。研究表明，光生物调节（PBM）可以改变巨噬细胞的极化状态并减轻炎症，尽管其细胞机制尚未完全阐明。在此，本研究探讨了PBM（980nm）对未分化巨噬细胞和M1型巨噬细胞的影响及其潜在机制。将RAW264.7细胞在不同激光参数（0.5、5.0和10.0J/cm）下进行激光照射，同时添加或不添加LY294002（PI3K途径抑制剂）。然后，使用共聚焦激光显微镜观察细胞分化；进行qPCR检测基因表达，使用蛋白质印迹法检测PI3K/AKT/mTOR途径中的蛋白质和活化的巨噬细胞标志物。所得结果表明，980nm PBM以剂量依赖性方式增加了M0型巨噬细胞中iNOS、Il-10、Arg1和Il-12以及炎性细胞因子Tnfα、IL-1β和Il-6的mRNA表达。更有趣的是，5J/cm的PBM降低了M1型巨噬细胞中iNOS、Il-12、Tnfα、IL-1β和Il-6的mRNA表达，并增加了Arg1和Il-10的表达，同时AKT和mTOR的磷酸化表达升高。此外，LY294002处理显著减弱了PBM诱导的M1型巨噬细胞极化。这些结果表明，980nm PBM可通过PI3K/AKT/mTOR途径激活M0型巨噬细胞并增加M2/M1比率。
