重复烟雾吸入建立肺损伤大鼠模型及肺纤维化评估

李虎明; 纵加强; 陈旭昕; 丁毅伟; 韩志海

doi:10.3969/j.issn.2095-5227.2022.08.009

重复烟雾吸入建立肺损伤大鼠模型及肺纤维化评估

Establishment of lung injured rat model and evaluation of lung fibrosis caused by repeated smoke inhalation

摘要

摘要:
背景烟雾吸入性损伤后期研究较少，且后期是否发生肺纤维化仍存在争议，建立不同烟雾吸入性肺损伤动物模型，评估损伤后期肺纤维情况，对研究烟雾吸入致伤机制有重要意义。
目的建立重复烟雾吸入性肺损伤大鼠模型，探讨其肺纤维化情况。
方法将健康成年雄性SD大鼠随机分为4组，每组30只。其中A组为空白对照，B组烟雾吸入1次(1次/d × 1 d，15 min/次)，C组烟雾吸入3次(1次/d × 3 d，15 min/次)，D组烟雾吸入7次(1次/d × 7 d，15 min/次)。烟雾吸入各组分别于吸入完成后第1天、第7天和第28天采集大鼠肺组织标本，检测炎性指标、血气分析、湿干比和肺组织病理染色。
结果 A组大鼠无死亡，B、C和D组烟雾吸入后分别死亡4只、7只和10只。与A组相比，B组第1天肺组织湿干比升高，白细胞介素-6和肿瘤坏死因子升高，碳氧血红蛋白(carboxyhemoglobin，COHb)升高，动脉血氧分压(PaO₂)下降。B组病理染色示肺泡结构弥漫性破坏，肺泡腔内可见红细胞、白细胞浸润，肺泡间隔增厚，炎细胞浸润明显，第7天时各指标较第1天好转，第28天部分指标恢复正常，Masson染色无明显胶原纤维沉积。与B组相比，C组和D组湿干比、炎性指标、COHb有升高趋势，特别是第28天Masson染色出现明显蓝色胶原纤维沉积。
结论重复烟雾吸入可以导致肺组织更明显的炎性反应，肺损伤程度增加，特别是后期可出现更明显的肺组织纤维化。

Abstract:
Background There are few studies on damage after smoke inhalation injury and it is still controversial about lung fibrosis after smoke inhalation injury in the advanced stage. Establishment of an animal model of smoke inhalation and evaluation of the lung fibrosis are of great significance to investigate the injury mechanism caused by smoke inhalation.
Objective To establish lung injured rat model after repeated smoke inhalation and explore the lung injured status.
Methods A total of 120 healthy male SD rats were divided into 4 groups at random, with 30 rats in each group, group A as control group without any intervention, group B was exposed to smoke inhalation for once (15min each time per day for one day), group C was exposed to smoke inhalation for three times (15min each time per day for three days), group D was exposed to smoke inhalation for seven times (15min each time per day for seven days). Rat lung tissues were obtained on the 1^st day, the 7^th day and the 28^th day among each group for inflammation test, arterial blood gas analysis, wet to dry ratio calculation and pathological staining.
Results There was no death in the group A, however, 4, 7, and 10 rats died in groups B, C, and D, respectively. Compared with the group A, wet to dry ratio, IL-6, TNF-α, and COHb increased in the group B on the first day, while the PaO₂ of arterial blood decreased. Pathology results showed pulmonary alveoli was destroyed at mass, the alveolar cavity was infiltrated with red and white blood cells, and the alveolar septum got thicker and the infiltration of inflammatory cells was obvious. On the 7^th day, the indexes in the group B were partly improved compared with that of the 1^st day; On the 28^th day, some parameters returned to normal and the Masson staining showed no manifest collagen fiber deposition. Compared with the group B, the wet to dry ratio, inflammation index and COHb in group C and group D had an increasing trend, and the Masson staining on the 28^th day showed obvious blue collagen fiber desposition.
Conclusion Repeated smoke inhalation may induce more serious inflammation reaction in lung tissue and aggravate the injury of lung, and more obvious pulmonary fibrosis may occur in the later stage.

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