The dynamic changes of collagen fiber in quartz-induced pulmonary fibrosis in mice with picrosirius red plus polarized microscopy in vivo

目的探讨天狼猩红-偏振光法结合定量图像分析观察石英引起肺纤维化过程中胶原纤维动态变化的应用价值.方法采用暴露式气管内注入法,对小鼠一次性染石英(0.2g/kg).染石英后第1、3、5、7、14、28天各处死8只小鼠,以苦味酸天狼猩红-偏振光结合定量图像分析观察其肺组织Ⅰ型胶原、Ⅲ型胶原的分布及变化,用氯胺T法测定肺组织羟脯氨酸含量.结果随染石英后时间的延长,小鼠肺两种胶原含量均呈增多趋势,早期Ⅲ型胶原增多较Ⅰ型胶原明显,而到后期,则以Ⅰ型胶原增加为主;Ⅰ/Ⅲ型胶原比值随着染石英时间延长呈现升高趋势,到第28天(1.49±0.39)与对照组(0.59±0.24)的差异出现统计学意义(P＜0.05);胶原总面积与肺组织羟脯氨酸含量间呈现正相关(r2=0.928 5,P＜0.01).结论天狼猩红-偏振光法结合定量图像分析是反映矽肺肺纤维化过程中胶原动态变化的一种准确、可行的方法.To investigate the use value of picrosirius red staining plus polarized microscopy to observe the dynamic changes of collagen fiber in lung fibrosis in silicotic mice model.The experimental mice were divided into control and quartz groups. 0.2 g/kg weight of quartz was injected intratracheally in quartz group. Lung tissues were collected at the 1st, 3rd, 5th, 7th, 14th and 28th day after injection respectively. Lung tissue slides were stained with picrosirius red. With the aid of polarized microscope, image analysis software, the distribution and change of type I and type III collagen could be qualitatively and quantitatively analyzed. Lung tissue hydroxyproline was determined by chloramines T method.In early stage the predominant increment was type III collagen, but in late stage type I was predominant. The contents of both type collagen tended to increase as postexposure time prolonged. The time course of the ratio of type I to type III showed increasing trend, and there was a statistical significance on day 28 (1.49 +/- 0.39 vs 0.59 +/- 0.24, P < 0.05). The total area of collagen was positively correlated with hydroxyproline concentration of lung tissue (r(2) = 0.928 5, P < 0.01).Picrosirius red staining combined with polarized microscopy and digital image processing is a useful method to elucidate collagen accumulation, distribution and subtype ratio in silicosis.PubMed中文核心期刊要目总览(PKU)中国科学引文数据库(CSCD)05361-3632