两种肺纤维化模型小鼠肺组织胶原含量定量分析方法的比较研究

A comparative study on two quantitative analysis methods for pulmonary collagen in fibrosis-sensitive and fibrosis-resistant mice

目的利用两种肺纤维化小鼠模型,对肺组织中胶原表达定量分析的两种方法——天狼猩红染色图像分析和羟脯氨酸测定法进行比较。方法采用20 Gy60Coγ射线胸部局部照射C57BL/6J和C3H/HeN两种小鼠分别建立敏感性/抗性肺纤维化模型,照射后3 m活杀取肺组织,左肺连续切片,根据均匀系统随机抽样的原则,在连续切片中均匀随机抽取一组切片,进行天狼猩红染色及图像分析;右肺采用碱水解法进行羟脯氨酸含量测定。结果两种分析方法结果均表明,γ射线照射后3 m C57BL/6J小鼠肺组织胶原含量较照射前明显增高,且显著高于C3H/HeN小鼠肺组织中胶原含量。天狼猩红染色图像分析法显示,C57BL/6J小鼠肺组织胶原面密度为C3H/HeN小鼠的2.29倍,而羟脯氨酸测定法显示,C57BL/6J小鼠肺组织羟脯氨酸含量为C3H/HeN小鼠的1.34倍。结论天狼猩红染色图像分析法较羟脯氨酸含量测定法能较理想地反映肺泡壁胶原含量的变化,且能更客观反映敏感性/抗性肺纤维化小鼠胶原表达的差异。

Objective To compare two quantitative methods, namely picrosirius staining and image analysis and hydroxyproline mensuration, used for pulmonary collagen analysis in fibrosis-sensitive and fibrosis-resistant mice. Methods C57BL/6J and C3H/HeN mice irradiated by 20 Gy 60γCo gamma rays were used as animal models. 3 months after irradiation, mice were sacrificed. Left lungs were embedded in paraffin blocks, then serial sections with thickness of 6μm were cut. Ten sections were uniformly and randomly sampled from the consecutive sections according to the principles of systematic, uniform and random sampling. One series of sections were stained with picrosirius and then image analysis was done. Hydroxyproline mensuration by alkali hydrolyzation was used to analyze the content of hydroxyproline in right lung. Results By these two quantitative analysis methods, significant increase of collagen deposi- tion were found in lung of C57BL/6J mice 3 months after irradiation and the pulmonary collagen contents of C57BL/6J mice was more than those of C3H/HeN mice. The area density of pulmonary collagen in C57BL/6J mice was 2.29 times that of C3H/HeN mice of pulmonary hydroxyproline in C57BL/6J mice was hydroxyproline mensuration. Conclusions showed by picrosirius staining, while the content 1.34 times that of C3H/HeN mice detected by Picrosirius staining and image analysis is more satisfactory than hydroxyproline mensuration for detecting change of collagen in alveolar wall of two animal models