630 nm LED照射对小鼠巨噬细胞向M1型极化的抑制作用

Inhibitory effect of 630 nm LED irradiation on the polarization of murine M1-type macrophages

目的:探讨630 nm发光二极管(LED)照射对体内外小鼠巨噬细胞向M1型极化及功能的影响。方法:以小鼠巨噬细胞系RAW264.7细胞和原代腹腔巨噬细胞为实验对象,在脂多糖(LPS)和干扰素-γ(IFN-γ)诱导其向M1型极化前、后用光照功率密度为8 mW/cm^(2)的630 nm LED照射细胞,利用CCK-8法测定细胞存活率,流式细胞术检测细胞表面活化/共刺激分子CD80和CD86的表达水平,实时荧光定量PCR(RT-qPCR)和/或酶联免疫吸附分析(ELISA)检测肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和白细胞介素-1β(IL-1β)等的表达与分泌。结果:630 nm LED照射对RAW264.7细胞的存活率无显著影响,但可显著降低经LPS和IFN-γ极化后RAW264.7细胞的CD80、CD86蛋白表达水平及促炎细胞因子TNF-α、IL-6和IL-1β的mRNA表达量(P<0.05)。体内外630 nm LED照射可抑制在体外经LPS和IFN-γ刺激极化后小鼠原代腹腔巨噬细胞TNF-α、IL-6的分泌水平(P<0.05)。结论:630 nm LED照射可抑制巨噬细胞向M1型极化和促炎因子的表达与分泌,这可为M1型巨噬细胞参与的炎症性疾病的治疗提供一些新的思路。

Objective:To investigate the effect of 630 nm LED irradiation on the polarization and function of murine M1-macrophages both in vitro and in vivo.Methods:Murine macrophage cell line RAW264.7 and primary peritoneal macrophages were irradiated with 630 nm LED(at a power density of 8 mW/cm^(2))either before or after the induction of M1-polarization by lipopolysaccharide(LPS)and interferon-γ(IFN-γ).Cell viabilities were measured by CCK-8 assay.Levels of surface activation/costimulatory molecules CD80&CD86 were determined by flow cytometry,and expressions of proinflammatory cytokines including tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)&interleukin-1β(IL-1β)were analyzed by quantitative real-time PCR(RT-qPCR)and/or enzyme-linked immunosorbent assay(ELISA).Results:Despite no effect on cell viabilities,630 nm LED irradiation,either before or after the induction of M1-polarization by LPS and IFN-γ,not only significantly reduced surface expressions of CD80 and CD86 on RAW264.7 cells,but also hampered their mRNA exprssion and productions of pro-inflammatory cytokines TNF-α,IL-6 and IL-1β(P<0.05).Moreover,pretreating mice with 630 nm LED followed by in vitro LED irradiation of cells repressed the ability of primary murine peritoneal macrophages to secrete TNF-αand IL-6 upon activation by LPS and IFN-γin vitro as well(P<0.05).Conclusion:630 nm LED irradiation blunts the M1 polarization and pro-inflammatory cytokine production of murine macrophages,which may provide new therapeutic strategy for the clinical treatment of M1 macrophage-mediated inflammatory diseases.