中国呼吸与危重监护杂志

中国呼吸与危重监护杂志

建立小鼠肺纤维化合并肺气肿模型的实验研究

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目的研究采用细颗粒物(PM2.5)经鼻滴入联合臭氧(O3)吸入建立小鼠肺纤维化合并肺气肿(CPFE)模型的方法的可行性,为 CPFE 模型的建立提供参考。方法雄性 C57/BL6 小鼠随机分为磷酸盐缓冲液(PBS)滴入+空气吸入组、PBS 滴入+O3 吸入组和 PM2.5 滴入+O3 吸入组,每组 8 只。小鼠接受经鼻滴入 PBS 或 PM2.5 混悬液(7.8 mg/kg),24 h 后接受吸入空气或 O3,每周两次,连续 8 周。8 周后,检测肺功能、支气管肺泡灌洗液(BALF)细胞计数和分类、苏木精-伊红染色观察肺组织病理形态学变化,包括炎症积分、平均肺泡内衬间隔(Lm),Masson 染色观察测定气道上皮下胶原沉积厚度(SEc/Pbm),同时测定肺组织羟脯氨酸含量。结果与 PBS 滴入+空气吸入组相比,PBS 滴入+O3 吸入组的吸气量(IC)增加,肺总量(TLC)增加,肺顺应性(Cchord)增加,FEV25(第 25 ms 的用力呼气量)/FVC(用力肺活量)降低,BALF 细胞总数增加,Lm 增大,肺组织炎症积分增加,SEc/Pbm 与羟脯氨酸无变化;PM2.5 滴入+O3 吸入组的 IC 降低,功能残气量(FRC)增加,TLC 增加,Cchord 降低,FEV25/FVC 与 FEV50(第 50 ms 的用力呼气量)/FVC 降低,BALF 细胞总数增加,肺组织炎症积分增加,Lm 增大,SEc/Pbm 和羟脯氨酸含量增加。与 PBS 滴入+O3 吸入组相比,PM2.5 滴入+O3 吸入组的 IC 降低,FRC 增加,Cchord 降低,FEV25/FVC 与 FEV50/FVC 降低,BALF 细胞总数增加,肺组织炎症积分增加,SEc/Pbm 和羟脯氨酸含量增加。结论对小鼠进行 8 周的 PM2.5 经鼻滴入联合 O3 吸入可成功建立 CPFE 模型。

ObjectiveTo study the possbility of using intranasal instillation of fine particulate matter (PM2.5) combined with inhalation of ozone (O3) to establish mouse model of combined pulmonary fibrosis and emphysema (CPFE), and to provide a reference for the establishment of CPFE model.MethodsMale C57/BL6 mice were divided randomly into phosphate buffer saline (PBS) intranasal instillation+air inhalation group, PBS intranasal instillation+O3 inhalation group and PM2.5 intranasal instillation+O3 inhalation group, with 8 mice in each group. The mice were intranasally instilled with PBS or PM2.5 suspension (7.8 mg/kg) followed by air or ozone inhalation 24 hours later, twice a week over 8 weeks. Lung function, bronchoalveolar lavage fluid (BALF) cell counts and classification were detected, the pathological changes of lung tissues in hematoxylin-eosin staining were observed, including inflammation scores and mean linear intercept (Lm). The thickness of collagen deposition in subepithelium was measured in lung tissues in Masson staining, and simultaneously hydroxyproline contents in lung tissues were determined.ResultsCompared to PBS instillation+air inhalation group, inspiratory capacity (IC), total lung capacity (TLC) and chord compliance (Cchord) were increased, FEV25 (the forced expiratory volume at 25 ms)/FVC (forced vital capacity) was decreased, total cell counts in BALF, Lm and lung inflammatory scores were increased, the thickness of the subepithelial collagen layer (SEc/Pbm) or hydroxyproline contents was not changed in PBS instillation +O3 inhalation group; IC was decreased, functional residual capacity (FRC) was increased, TLC was increased, Cchord was decreased, FEV25/FVC and FEV50 (the forced expiratory volume at 50 ms)/FVC were decreased, total cell counts in BALF, Lm, lung inflammatory scores, SEc/Pbm and hydroxyproline contents were increased in PM2.5 instillation+O3 inhalation group. Compared to PBS instillation+O3 inhalation group, IC was decreased, FRC was increased, Cchord was decreased, FEV25/FVC and FEV50/FVC were decreased, total cell counts in BALF, Lm, lung inflammatory scores, SEc/Pbm and hydroxyproline contents were increased in PM2.5 instillation +O3 inhalation group.ConclusionCPFE mouse model can be successfully established by PM2.5 intranasal instillation combined with ozone inhalation for consecutive 8 weeks.

关键词: 细颗粒物; 臭氧; 小鼠模型; 肺纤维化合并肺气肿

Key words: Fine particulate matter; Ozone; Mouse model; Combined pulmonary fibrosis and emphysema

引用本文: 李锋, 徐蒙蒙, 王沐昀, 陈宇清, 张海, 张妍蓓. 建立小鼠肺纤维化合并肺气肿模型的实验研究. 中国呼吸与危重监护杂志, 2018, 17(6): 604-608. doi: 10.7507/1671-6205.201806007 复制

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