吡非尼酮联合尼达尼布治疗小鼠肺纤维化的效果

The effect of Pirfenidone combined with Nintedanib treating pulmonary fibrosis in mice

目的探讨吡非尼酮联合尼达尼布联合治疗肺纤维化的效果和方法。方法选取90只清洁级6～8周龄雄性昆明小鼠,适应性饲养1周后,随机取10只作为对照组,其余80只用博来霉素制备肺纤维化模型,用Buxco动物肺功能分析系统测定小鼠的肺功能,Penh值上升至1.50以上显示造模成功。从造模成功的小鼠中随机抽取40只,随机分为对照组(0.1 ml生理盐水,每天3次)、吡非尼酮组(400 mg/kg,每天3次)、尼达尼布组(150 mg/kg,每天2次)、吡非尼酮(400 mg/kg,每天3次)联合尼达尼布组(150 mg/kg,每天2次),连续给药3周。上法测定肺功能的Penh值,然后将小鼠麻醉后处死,收集肺泡灌洗液,离心获得取细胞,采用瑞士-姬姆萨染色后,在倒置显微镜下计数小鼠肺泡灌洗液中的白细胞。结果 80只小鼠造模,50只小鼠(62.5%)造模成功。治疗后,吡非尼酮联合尼达尼布组的肺功能Penh值小于吡非尼酮组(P<0.05)、尼达尼布组(P<0.05)和对照组(P<0.01)。吡非尼酮联合尼达尼布组的白细胞计数少于吡非尼酮组(P<0.05)、尼达尼布组(P<0.05)和对照组(P<0.01)。结论吡非尼酮联合尼达尼布的治疗效果最佳,为临床使用该方法治疗肺纤维化奠定了基础。

Objective To explore the effect and method of Pirfenidone combined with Nintedanib treating pulmonary fibrosis in mice. Methods All of 90 male Kunming mice aged 6-8 weeks were selected, After 1 week of adaptive feeding, and among them, 10 mice were randomly selected as control group, and the other 80 mice were treated with Bleomycin to establish pulmonary fibrosis model. The pulmonary function of the mice was measured by Buxco animal pulmonary function analysis system. The Penh value increased to more than 1.5, and the model was established successfully. 40 model mice were selected randomly from successful mice modeling and randomly divided into control group (0.1 ml normal saline, 3 times a day), Pirfenidone group (400 mg/kg, 3 times a day), Nintedanib group (150 mg/kg, 2 times a day), Pirfenidone (400 mg/kg, 3 times a day) combined with Nintedanib group (150 mg/kg, 2 times a day). The mice were gavaged for 3 weeks. Penh value was measured using the above-mentioned method. Then the mice were sacrificed after anaesthetized. The alveolar lavage fluid was collected and centrifuged, then the cells were obtained. After Swiss-Giemsa staining, the white cells in alveolar lavage fluid of mice were counted under inverted microscope. Results A total of 80 mice were modeled and 50 mice (62.5%) were successfully modeled. After treatment, the Penh value in Pirfenidone group combined with Nintedanib group were lower than those in Pirfenidone group (P<0.05), Nintedanib group (P<0.05) and control group (P<0.01). The leukocyte of Pirfenidone combined with Nintedanib group was fewer than that of Pirfenidone group (P<0.05), Nintedanib group (P<0.05) and control group (P<0.01). Conclusion The therapeutic effect of Pirfenidone combined with Nintedanib is the best, which lays a foundation for the clinical use of this method in the treatment of pulmonary fibrosis.