630 nm LED红光通过AKT/FOXO3a信号通路抑制人巨噬细胞炎症因子释放的研究  

Study of 630 nm LED irradiation inhibits human macrophage inflammatory factor releases through AKT/FOXO3a pathway

作　　者：潘昱霖 林默楠 张凯波 曹国丁 易斌 何军 刘海亮 张凤民[1] PAN Yulin;LIN Monan;ZHANG Kaibo;CAO Guoding;YI Bin;HE Jun;LIU Hailiang;ZHANG Fengmin(Department of Microbiology,Wu Lien-Teh Institute,Harbin Medical University,the Heilongjiang Key Laboratory of Immunity and Infection,Harbin 150081,China;Beijing Truwin Optoelectronic Medical,Beijing 100176,China)

机构地区：[1]哈尔滨医科大学医学微生物学教研室、伍连德研究所、黑龙江省感染与免疫重点实验室,哈尔滨150081 [2]北京创盈光电医疗科技公司,北京100176

出　　处：《国际免疫学杂志》2024年第2期117-123,共7页International Journal of Immunology

基　　金：国家重点研发计划资助项目(2017YFB0403805);黑龙江省自然科学基金优秀青年项目(YQ2020H004)。

摘　　要：目的探讨630 nm发光二极管(light emitting diode,LED)红光照射对人外周血单核细胞系THP-1来源巨噬细胞炎症反应的作用,寻找其可能发挥作用的信号通路与分子机制。方法人外周血单核细胞THP-1,经佛波酯(phorbol myristate acetate,PMA)诱导为THP-1来源巨噬细胞,并用脂多糖(lipopolysaccharide,LPS)处理制备巨噬细胞炎症模型;未经630 nm LED红光照射的细胞设为对照组,实验组各组细胞接受光照能量强度分别为14.4、28.8和43.2 J/cm^(2);对不同能量强度LED红光照射处理后的巨噬细胞进行实时荧光定量PCR(real-time reverse transcription polymerase chain reaction,RT-qPCR)和酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA),检测炎症因子白细胞介素(interleukin,IL)-1β和肿瘤坏死因子(tumor necrosis factor,TNF)-α的mRNA与蛋白表达,Western blot检测蛋白激酶B(protein kinase B,AKT)和叉头框蛋白O3a(forkhead box O3a,FOXO3a)蛋白及其磷酸化(p-AKT、p-FOXO3a)表达,并分析其分子通路。结果巨噬细胞经LPS诱导后,与对照组相比,实验组细胞中iNOS的mRNA表达显著升高,同时伴随炎症因子IL-1β、TNF-α的mNRA表达水平显著提升,差异有统计学意义(t值分别为-48.73、-80.96、-38.45,P值均<0.001)。M1巨噬细胞经630 nm LED红光照射处理后,细胞中炎症因子IL-1β、TNF-α的mRNA表达水平明显降低,同时IL-1β、TNF-α的蛋白表达水平下降,p-AKT/AKT蛋白比值和p-FOXO3a/FOXO3a蛋白比值明显降低,差异有统计学意义(t值分别为60.18、25.30、14.79、17.31、23.64、87.50,P值均<0.05)。SC79干预后,M1巨噬细胞的p-AKT/AKT蛋白比值显著升高,IL-1β的mRNA、蛋白表达显著增加,差异有统计学意义(t值分别为-18.08、-26.43、-18.06,P值均<0.05)。SC79激活M1巨噬细胞AKT磷酸化后,630 nm LED红光可以明显降低SC79干预后M1巨噬细胞中p-AKT/AKT蛋白比值,同时明显下调IL-1β、TNF-α的mRNA表达和IL-1β、TNF-α的蛋白表达,差异有统计学意义Objective To investigate the effect of 630 nm light emitting diode(LED)red light irradiation on the inflammatory response of human peripheral blood mononuclear cells THP-1-derived macrophages,and its potential signaling pathways and molecular mechanisms.Methods Human peripheral blood mononuclear cells,THP-1,were induced into THP-1-derived macrophages by phorbol myristate acetate(PMA)and treated with lipopolysaccharide(LPS)to prepare a macrophage inflammation model.The cells not irradiated with 630 nm LED red light were set as the control group,the cells in each experimental group received light energy intensities of 14.4,28.8 and 43.2 J/cm^(2).Real-time reverse transcription polymerase chain reaction(RT-qPCR)and enzyme linked immunosorbent assay(ELISA)were performed to detect the mRNA and protein expression of inflammatory factors interleukin(IL)-1βand tumor necrosis factor(TNF)-αin macrophages after treatment with LED red light irradiation of different energy intensities,and Western blot to detect the protein expression of kinase B(AKT)and forkhead box O3a(FOXO3a)protein and their phosphorylation(p-AKT,p-FOXO3a),and their molecular pathways were analyzed.Results After macrophages were induced by LPS,the mRNA expression of iNOS in the cells of the experimental group was significantly elevated compared with the control group,accompanied by a significant increase in the mRNA expression level of the inflammatory factors IL-1βand TNF-α,and the difference was statistically significant(t values were-48.73,-80.96,-38.45,respectively,all P values<0.001).After M1 macrophages were treated with 630 nm LED red light irradiation,the mRNA expression levels of the inflammatory factors IL-1βand TNF-αin the cells were significantly reduced,along with the protein expression levels of IL-1βand TNF-α,and the p-AKT/AKT protein ratios and p-FOXO3a/FOXO3a protein ratios were significantly reduced,with statistically significant differences(t values were 60.18,25.30,14.79,17.31,23.64,87.50,respectively,all P values<0.05).After SC

关 键 词：630 nm LED红光 巨噬细胞 蛋白激酶B 叉头蛋白O3a 

分 类 号：R392[医药卫生—免疫学]