小动物X射线辐照仪与小鼠放射性肺纤维化模型的构建  

The small animal X-ray irradiators and the establishment of the mice model of radiation-induced pulmonary fibrosis

作　　者：章登峰 李科君 徐畅[1] 杜利清[1] Zhang Dengfeng;Li Kejun;Xu Chang;Du Liqing(Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine,Institute of Radiation Medicine,Chinese Academy of Medical Sciences,Peking Union Medical College,Tianjin 300192,China)

机构地区：[1]中国医学科学院北京协和医学院放射医学研究所,天津市放射医学与分子核医学重点实验室,天津300192

出　　处：《国际放射医学核医学杂志》2024年第10期623-630,共8页International Journal of Radiation Medicine and Nuclear Medicine

摘　　要：目的探讨采用小动物X射线辐照仪构建小鼠放射性肺纤维化(RIPF)模型的方法。方法将24只C57BL/6J小鼠随机分为4组(对照组、20 Gy照射组、22 Gy照射组和24 Gy照射组),每组6只,3个照射组使用小动物X射线辐照仪进行单次右肺照射,于照射后120 d取材。观察小鼠照射后的体重、右肺组织经苏木精-伊红和马松三色染色后的肺部组织病理学改变;采用Western blot和逆转录实时定量聚合酶链式反应检测小鼠肺组织纤维黏连蛋白1(FN1)和α-平滑肌肌动蛋白(α-SMA)纤维化相关蛋白的表达;检测肺组织中羟脯氨酸含量和转化生长因子β1(TGF-β1)信号通路的变化。计量资料的组间比较采用单因素方差分析。结果对照组小鼠照射2周后的体重为(28.8±3.3)g,20、22、24 Gy照射组小鼠的体重分别为(26.2±4.6)、(26.4±3.3)、(26.6±3.3)g,差异无统计学意义(F=0.654,P>0.05)。与对照组相比,照射组小鼠右肺组织出现肺泡结构破坏,肺泡间隔增厚,伴有明显的胶原纤维沉积。照射组小鼠肺组织中FN1和α-SMA纤维化相关蛋白的表达水平较对照组明显升高,24 Gy照射组小鼠FN1基因的转录水平(2-ΔΔCt:2.1±0.6)明显高于对照组(2-ΔΔCt:0.6±0.1),20 Gy照射组小鼠α-SMA基因的转录水平(2^(-ΔΔCt):2.1±0.7)明显高于对照组(2-ΔΔCt:1.1±0.3),差异均有统计学意义(F=12.470、4.320,均P<0.05)。对照组小鼠右肺组织的羟脯氨酸含量为(500.2±17.1)μg/mg,20、22、24 Gy照射组小鼠右肺组织的羟脯氨酸含量分别为(1199.7±159.2)、(1357.1±36.8)、(1649.3±213.4)μg/mg。与对照组相比,不同剂量X射线照射组小鼠右肺组织的羟脯氨酸含量均明显增加,且差异有统计学意义(F=13.170,P<0.001),并伴有TGF-β1信号通路的激活。以上结果表明小鼠RIPF模型构建成功。结论采用小动物X射线辐照仪进行单次右肺照射(剂量20~24 Gy)120 d后小鼠出现明显肺纤维化,可成功构建小鼠RIPF模型。Objective To establish a mice model of radiation-induced pulmonary fibrosis(RIPF)with small-animal X-ray irradiators.Methods Twenty-four C57BL/6J mice were randomly classified into four groups.One group received sham irradiation(control group,n=6),and the other three groups were irradiated with a single dose of X-ray in the right lung by small-animal X-ray irradiators(20 Gy irradiation group,n=6;22 Gy irradiation group,n=6;24 Gy irradiation group,n=6).The mice were sacrificed 120 days after irradiation and lung tissues were extracted.Observe the body weight,the pathological changes in the right lung tissue of mice after irradiation stained with hematoxylin-eosin and Masson′s trichrome;Western blot and real-time quantitative polymerase chain reaction were used to detect the expression of fibronectin 1(FN1)andα-smooth muscle actin(α-SMA)fibrosis related proteins in mice lung tissue.Detecting changes in hydroxyproline contents,and transforming growth factor beta 1(TGF-β1)signaling pathway in lung tissue.Quantitative data were analyzed by one-way ANOVA.Results At two weeks after irradiation,mice in the control group had a body weight of(28.8±3.3)g.Mice in the 20,22,and 24 Gy irradiation groups had body weights of(26.2±4.6),(26.4±3.3),and(26.6±3.3)g,respectively.The difference was not statistically significant(F=0.654,P>0.05).Compared with the control group,the right lung tissue of the irradiated groups mice showed structural damage to the alveoli,thickening of the alveolar septa,and significant deposition of collagen fibers.The expression levels of fibrosis related proteins,FN1,andα-SMA significantly increased in the lungs from radiation-injured groups.The transcription levels of FN1 in the 24 Gy irradiation group was significantly higher than those in the control group and the difference was statistically significant(2^(-ΔΔCt):(2.1±0.6)vs.(0.6±0.1),F=12.470,P<0.05).The transcription levels ofα-SMA in the 20 Gy irradiation group were significantly higher than those in the control group and the differen

关 键 词：放射性 肺纤维化 C57BL小鼠 动物模型 小动物X射线辐照仪 

分 类 号：R818.8[医药卫生—放射医学]