血小板反应蛋白-1敲除对放射性肺纤维化的影响及其机制  

Effect and underlying mechanism of thrombospondin-1 knockout on radiationinduced pulmonary fibrosis in mice

作　　者：焦露 李杰[1] 张静 陈用来 郝玉徽[1] QIAO Lu;LI Jie;ZHANG Jing;CHEN Yonglai;HAO Yuhui(Institute of Combined Injury,Faculty of Military Preventive Medicine,Army Medical University(Third Military Medical University),Chongqing,400038,China)

机构地区：[1]陆军军医大学(第三军医大学)军事预防医学系复合伤研究所,重庆400038

出　　处：《陆军军医大学学报》2023年第9期975-981,共7页Journal of Army Medical University

基　　金：重庆市自然科学基金面上项目(2022NSCQ-MSX4675)。

摘　　要：目的探讨血小板反应蛋白-1(thrombospondin-1,TSP-1)对放射性肺纤维化的影响及其可能机制。方法选取6~8周龄雄性野生型C57BL/6小鼠(wild type,WT)和全身性敲除TSP-1小鼠(TSP-1^(-/-)),将小鼠分为野生辐照组(WT+IR)和TSP-1敲除辐照组(TSP-1^(-/-)+IR),每组各6只。采用60 Coγ射线全肺照射16 Gy建立放射性肺纤维化小鼠模型。照射后第16周,HE染色观察小鼠肺组织形态变化,天狼猩红染色观察肺组织胶原表达水平,qRT-PCR法检测小鼠肺组织中TGF-β1和CollagenⅠ的mRNA表达水平,micro-CT评价肺损伤和肺实变情况,FlexiVent系统评价小鼠肺功能水平,免疫荧光染色检测肺组织LC-3和p62的蛋白表达,透射电镜观察肺组织细胞自噬体。结果与野生辐照组相比,TSP-1敲除辐照组小鼠肺损伤减轻,肺组织胶原沉积减少,TGF-β1和CollagenⅠ的mRNA表达水平显著降低(P<0.01),肺功能水平有所改善。免疫荧光和透射电镜结果显示TSP-1敲除辐照组小鼠肺组织出现细胞自噬。结论TSP-1敲除能够减轻放射性肺纤维化,其机制可能与细胞自噬激活有关。Objective To investigate the effect of thrombospondin-1(TSP-1)knockout on radiationinduced pulmonary fibrosis and explore its underlying mechanism.Methods Male wild type C57BL/6 mice(6~8 weeks old,WT mice)and TSP-1 knockout mice(TSP-1-/-mice)were subjected and received 16 Gy 60 Coγ-ray whole lung irradiation(IR)to induce radiation pulmonary fibrosis,and then,the mice were assigned into WT+IR group and TSP-1-/-+IR group,respectively,with 6 mice in each group.In 16 weeks after irradiation,HE staining was performed to observe the morphological changes in lung tissues,and Sirius red staining was used to observe the collagen expression in lung tissues.qRT-PCR was employed to detect the mRNA expression of TGF-β1 and CollagenⅠin lung tissues.Micro-CT scanning was adopted to evaluate lung injury and substantial lung changes.FlexiVent system was employed to assess pulmonary function.Immunofluorescence staining was conducted to measure the protein levels of LC-3 and p62 in lung tissues.Transmission electron microscopy(TEM)was used to observe the cellular autophagic vesicles in lung tissues.Results Compared with the WT+IR group,the TSP-1^(-/-)+IR group had reduced lung injury and collagen deposition in lung tissue,significantly lower mRNA levels of TGF-β1 and CollagenⅠin lung tissue(P<0.01),and improved pulmonary function.Immunofluorescence assay and TEM showed that autophagy was observed in the mice of TSP-1^(-/-)+IR group.Conclusion TSP-1 knockout can alleviate radiation-induced pulmonary fibrosis,which may be related to activation of autophagy.

关 键 词：血小板反应蛋白-1 放射性肺纤维化 自噬 TGF-Β1 FlexiVent 

分 类 号：R341[医药卫生—基础医学] R563.9R818.023