NRP1调控M2型巨噬细胞极化介导电离辐射肺纤维化的作用研究

NRP1 regulates polarization-mediated ionizing radiation pulmonary fibrosis of M2 macrophages

目的:研究神经菌毛蛋白1(NRP1)对电离辐射肺损伤导致纤维化进程的影响,并探讨其与M2型巨噬细胞间的关系。方法:电离辐射肺纤维化(RIPF)组小鼠以6 Gy X射线全胸辐射建立电离辐射肺纤维模型;对照组即正常小鼠;EG00229给药组(EG00229为NRP1拮抗剂)小鼠辐射前7 d小鼠EG00229给药处理。HE、天狼星红、马松染色观察三组小鼠肺组织损伤和纤维化胶原沉积;ELISA检测三组小鼠肺泡灌洗液中TNF-α、IL-4、TGF-β1炎症因子水平;IHC和RT-PCR检测三组小鼠肺组织中Ⅰ型胶原(Col-Ⅰ)、α平滑肌肌动蛋白(α-SMA)和NRP1蛋白表达水平;免疫荧光检测三组小鼠肺组织中CD14和M2型巨噬细胞极化标志物精氨酸酶1(Arg1)、抵抗素样分子α(FIZZ1)、几丁质酶3样蛋白1(Ym1)蛋白表达水平。结果:HE染色显示RIPF组小鼠肺部呈现放射性肺纤维化病理形态表现,EG00229给药组小鼠肺组织纤维化损伤有明显改善;与对照组相比,RIPF组小鼠肺组织中Col-Ⅰ、α-SMA和NRP1及TNF-α、IL-4、TGF-β1和CD14表达水平显著升高,EG00229可显著抑制小鼠肺组织中Col-Ⅰ、α-SMA和NRP1以及TNF-α、IL-4、TGF-β1和CD14表达(P<0.05);与对照组相比,RIPF组小鼠肺组织中Arg1、FIZZ1和Ym1表达水平显著升高,EG00229可明显抑制小鼠肺组织中Arg1、FIZZ1和Ym1表达。结论:NRP1可能通过调控M2型巨噬细胞极化促进电离辐射肺纤维化的发生发展。

Objective:To study effect of Neurofibrin 1(NRP1)on fibrosis process caused by ionizing radiation lung injury,and to explore its relationship with M2 macrophages.Methods:Radiation-induced pulmonary fibrosis(RIPF)group mice were irradiated with 6 Gy X-ray whole chest to establish ionizing radiation lung fiber model;control group was normal mice;EG00229 administration group(EG00229 was NRP1 antagonist)mice were treatment with EG002297 days before irradiation.HE,Sirius red and Masson staining were used to observe lung tissue damage and fibrotic collagen deposition in three groups of mice;ELISA was used to detect levels of TNF-α,IL-4,TGF-β1 inflammatory factors in alveolar lavage fluid of three groups of mice;IHC and RT-PCR methods were used to detect protein expression levels of typeⅠcollagen(Col-Ⅰ),α-smooth muscle actin(α-SMA)and NRP1 in lung tissues of three groups of mice;immunofluorescence was used to detect lung tissues of three groups of mice CD14 and M2 macrophage polarization markers arginase 1(Arg1),resistin-like moleculeα(FIZZ1),chitinase 3(Ym1)protein expression levels.Results:HE staining showed that lungs of RIPF group showed radioactive pulmonary fibrosis pathological manifestations.Lung fibrosis injury of mice in EG00229 administration group was significantly improved;compared with control group,expression levels of Col-Ⅰ,α-SMA and NRP1 as well as TNF-α,IL-4,TGF-β1 and CD14 in lung tissues of mice in RIPF group were significantly increased.EG00229 could significantly inhibit expressions of Col-Ⅰ,α-SMA and NRP1 as well as TNF-α,IL-4,TGF-β1 and CD14 in mice lung tissue(P<0.05);compared with control group,expression levels of Arg1,FIZZ1 and Ym1 in lung tissue of RIPF group were significantly increased,and EG00229 could significantly inhibit expressions of Arg1,FIZZ1 and Ym1 in lung tissue of mice.Conclusion:NRP1 may promote occurrence and development of ionizing radiation pulmonary fibrosis by regulating polarization of M2 macrophages.