Li Kun, Liang Zhuowen, Zhang Jiawei, Zuo Xiaoshuang, Sun Jiakai, Zheng Qiao, Song Jiwei, Ding Tan, Hu Xueyu, Wang Zhe
Xijing Orthopaedics Hospital, Fourth Military Medical University, Xi'an, Shaanxi, China.
Lasers Med Sci. 2020 Sep;35(7):1509-1518. doi: 10.1007/s10103-019-02941-y. Epub 2020 Feb 17.
In spinal cord injury (SCI), inflammation is a major mediator of damage and loss of function and is regulated primarily by the bone marrow-derived macrophages (BMDMs). Photobiomodulation (PBM) or low-level light stimulation is known to have anti-inflammatory effects and has previously been used in the treatment of SCI, although its precise cellular mechanisms remain unclear. In the present study, the effect of PBM at 810 nm on classically activated BMDMs was evaluated to investigate the mechanisms underlying its anti-inflammatory effects. BMDMs were cultured and irradiated (810 nm, 2 mW/cm) following stimulation with lipopolysaccharide and interferon-γ. CCK-8 assay, 2',7'-dichlorofluorescein diacetate assay, and ELISA and western blot analysis were performed to measure cell viability, reactive oxygen species production, and inflammatory marker production, respectively. PBM irradiation of classically activated macrophages significantly increased the cell viability and inhibited reactive oxygen species generation. PBM suppressed the expression of a marker of classically activated macrophages, inducible nitric oxide synthase; decreased the mRNA expression and secretion of pro-inflammatory cytokines, tumor necrosis factor alpha, and interleukin-1 beta; and increased the secretion of monocyte chemotactic protein 1. Exposure to PBM likewise significantly reduced the expression and phosphorylation of NF-κB p65 in classically activated BMDMs. Taken together, these results suggest that PBM can successfully modulate inflammation and polarization in classically activated BMDMs. The present study provides a theoretical basis to support wider clinical application of PBM in the treatment of SCI.
在脊髓损伤(SCI)中,炎症是损伤和功能丧失的主要介质,主要由骨髓来源的巨噬细胞(BMDM)调节。光生物调节(PBM)或低水平光刺激已知具有抗炎作用,并且先前已用于SCI的治疗,尽管其确切的细胞机制仍不清楚。在本研究中,评估了810nm的PBM对经典激活的BMDM的影响,以研究其抗炎作用的潜在机制。在用脂多糖和干扰素-γ刺激后,培养并照射BMDM(810nm,2mW/cm)。分别进行CCK-8测定、2',7'-二氯荧光素二乙酸酯测定、ELISA和蛋白质印迹分析,以测量细胞活力、活性氧产生和炎症标志物产生。经典激活的巨噬细胞的PBM照射显著提高了细胞活力并抑制了活性氧的产生。PBM抑制了经典激活的巨噬细胞标志物诱导型一氧化氮合酶的表达;降低了促炎细胞因子肿瘤坏死因子α和白细胞介素-1β的mRNA表达和分泌;并增加了单核细胞趋化蛋白1的分泌。暴露于PBM同样显著降低了经典激活的BMDM中NF-κB p65的表达和磷酸化。综上所述,这些结果表明PBM可以成功调节经典激活的BMDM中的炎症和极化。本研究为支持PBM在SCI治疗中更广泛的临床应用提供了理论基础。