【摘要】 M1型巨噬细胞为经典活化型巨噬细胞,可分泌多种促炎因子和趋化因子促进炎症的发展。我们前期研究已经证明,630 nm发光二极管(LED)照射能抑制巨噬细胞释放炎症因子,然而,630 nm LED对巨噬细胞极化的调控作用及其分子机制尚不清楚。在本研究中,我们评估了630 nm LED照射对单核细胞(THP-1)来源的M1型巨噬细胞极化的影响,探讨了其对M1型巨噬细胞极化的作用机制。体外培养THP-1细胞,用佛波酯(PMA)将其诱导成巨噬细胞。用脂多糖(LPS)和干扰素-γ(IFN-γ)处理巨噬细胞,使其极化成M1型巨噬细胞,极化的同时用波长为630 nm、功率密度为40.02 mW/cm~2的LED对其进行照射。采用噻唑蓝(MTT)法、细胞计数试剂盒(CCK-8)法和乳酸脱氢酶(LDH)活性检测试剂盒检测细胞活力,用实时定量PCR法检测M1型巨噬细胞相关因子mRNA的表达水平,采用免疫印迹和细胞免疫荧光检测M1型巨噬细胞中STAT1的核转位。结果显示,M1型巨噬细胞经630 nm LED照射后,其活性未受影响,但M1型巨噬细胞相关因子干扰素调节因子5(IRF5)、趋化因子配体9(CXCL9)、CXL10、CXCL11 m RNA的表达水平显著降低,并且阻滞转录因子STAT1向核内转移。综上所述,630 nm LED照射可通过阻滞STAT1核转位抑制M1型巨噬细胞极化,此研究可能对治疗巨噬细胞极化相关疾病提供新的思路。更多还原

【Abstract】 M1 macrophages are classical activated macrophages, which can secrete a variety of pro-inflammatory factors and chemokines to promote the development of inflammation. Our previous studies have demonstrated that 630 nm light-emitting diode(LED) irradiation inhibits the release of inflammatory factors in macrophages. However, the effect of 630 nm LED on macrophage polarization and its molecular mechanism remains unclear. In this study, we evaluated the effect of 630 nm LED irradiation on the polarization of THP-1 derived M1 macrophages and explored the mechanism of its effect on the polarization of M1 macrophages. Human mononuclear cell line THP-1 was cultured in vitro and induced into macrophages by phorbol myristate acetate(PMA). Lipopolysaccharide(LPS) and interferon-γ(IFN-γ) treated cells were polarized into M1 macrophages,which were irradiated by LED with a wavelength of 630 nm and a power density of 40.02 mW/cm~2. CCK-8 colorimetry, MTT assay and lactate dehydrogenase(LDH) activity detection kit were used to detect cell viability. Real-time quantitative PCR was used to detect mRNA expression levels of related factors in M1 macrophages. Western blot and immunofluorescence were used to detect the nuclear translocation of STAT1 in M1 macrophages. The results showed that the cell viability of M1 macrophages did not alter after 630 nm LED irradiation. LED irradiation significantly reduced the mRNA expression levels of the interferon regulatory factor 5(IRF5) and C-X-C motif chemokine ligand(CXCL) 9, 10 and 11 and block the translocation of transcription factor STAT1. Taken together, 630 nm LED inhibits the polarization of M1 macrophages by blocked STAT1 nuclear translocation. This study may provide new therapeutic ideas for the treatment of diseases related to macrophage polarization.更多还原