不同剂量博来霉素滴鼻诱导C57BL/6 小鼠 肺纤维化模型的研究

Research on C57BL/6 mice model of pulmonary fibrosis induced by nasal instillation of different dose-bleomycin

摘要:

目的 探讨以鼻腔滴注博来霉素（BLM）的方式诱导C57BL/6 小鼠肺纤维化模型的最佳方法。 方法 7 ～ 8 周龄雄性小鼠30 只，分为0.10 u BLM 组、0.25 u BLM 组、0.40 u BLM 组，每组8 只，麻醉后分 别一次性滴鼻0.10、0.25 及0.40 u BLM ；对照组6 只，给予等体积的磷酸盐缓冲液。观察并记录小鼠体重变 化与生存率，实验结束后处死小鼠。用甲醛固定肺组织，苏木精- 伊红染色评价肺组织炎症变化；Masson 染 色评价肺纤维化变化。结果 给药后小鼠体重逐渐下降，并在第8 和9 天再次上升，第14 天逐渐趋近于对照组， 第28 天BLM 组所有小鼠肺部组织产生纤维化。各组肺纤维化评分比较，差异无统计学意义（P >0.05）。但 给药0.25 与0.40 u BLM 的小鼠死亡率高，个体间差异较大，不利于实验样本的收集与数据的统计分析。 结论 0.10 u BLM 滴鼻给药是复制C57BL/6 小鼠肺纤维化模型便捷、理想的实验方法。

Abstract:

Objective To explore the optimal methods of pulmonary fibrosis model induced by nasal instillation of bleomycin in C57BL/6 mice. Methods Thirty 7 to 8-week-old male mice were divided into four groups. Three experimental groups were respectively treated with different doses of 0.10 unit, 0.25 unit, 0.40 unit of bleomycin by nasal instillation after anesthesia (eight mice in each group). The control group were treated with the same volume of PBS (six mice in the group). The body weight and survival rate in each group were examined. At the end of the experiment, all of the mice were sacrificed, and we fixed the lung tissues in formalin, then cut them into slices. The extent of pulmonary inflammation was evaluated by HE staining. And the extent of pulmonary fibrosis was evaluated by Masson staining. Results The body weights of mouse declined gradually but increased on the 8 to 9th day. Until the 14th day, the body weights of drugs were nearly at the same level as the controls. At the 28th day, all of the mice got pulmonary fibrosis in BLM groups, and there was no significant statistical difference between pulmonary fibrosis scores of each group (P > 0.05). But the death rates were higher in groups of 0.25 unit and 0.4 unit, and there was a big difference in each individual. It was not good for the collection of experimental samples and statistical analysis of data. Conclusions Nasal instillation of 0.10 unit bleomycin is a convenient and ideal method in establishing C57BL/6mice model of pulmonary fibrosis.