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目的:探讨60Coγ射线照射对肺泡II型细胞和肺泡隔间质细胞的生物效应。方法:原代分离肺内II型上皮细胞(AT-Ⅱ)和间质细胞包括巨噬细胞和成纤维细胞,分别进行0、3、5、7Gy的γ射线照射,用细胞核嗜银染色观察照射对AT-Ⅱ增殖的影响;用酶谱分析检测照射后AT-Ⅱ和间质细胞培养上清中基质金属蛋白酶-2、-9(MMP-2,-9)的活性;用ELISA检测间质细胞培养上清中TGF-β1和IV型胶原含量。结果:AT-Ⅱ的核仁数量随照射剂量增加而增多,其中7Gy组最高;AT-II培养上清中MMP-2、-9活性随照射剂量增加呈先增高后降低趋势,间质细胞上清中MMP-2、-9活性和TGF-β1水平逐渐升高,IV型胶原分泌水平呈先降低后升高趋势。结论:放射性肺损伤早期,AT-Ⅱ、巨噬细胞和成纤维细胞均参与肺组织无效性重建过程,与晚期肺纤维化启动有一定的内在联系。
Objective: To investigate the biological effects of 60Coγ-ray irradiation on alveolar type II cells and alveolar septum cells. Methods: Primary isolated lung type II epithelial cells (AT-Ⅱ) and stromal cells including macrophages and fibroblasts were irradiated with gamma rays of 0, 3, 5, and 7 Gy, respectively. On the proliferation of AT-Ⅱ. The activity of matrix metalloproteinase-2, -9 (MMP-2, -9) in AT-Ⅱ and stromal cell culture supernatant was detected by zymography; TGF-β1 and type IV collagen content in cell culture supernatants. Results: The number of AT-II nucleolus increased with the increase of radiation dose, and the highest in 7Gy group. The activity of MMP-2 and -9 in AT-II culture supernatant increased first and then decreased with the increase of radiation dose. The serum levels of MMP-2, -9 and TGF-β1 gradually increased, and the type IV collagen secretion decreased first and then increased. Conclusion: In the early stage of radiation-induced lung injury, AT-Ⅱ, macrophages and fibroblasts are involved in the process of ineffective pulmonary remodeling, which is intrinsically associated with the initiation of advanced pulmonary fibrosis.