不同方式下气管内滴注脂多糖方法制作急性呼吸窘迫综合征大鼠模型

目的探讨内窥镜下辅助滴注脂多糖及气管暴露法滴注脂多糖诱导急性呼吸窘迫综合征（ARDS）大鼠动物模型的差异。 方法32只Sprague Dawley大鼠分为EC组（内窥镜下辅助气管插管滴注等渗NaCl溶液）、EL组（内窥镜下辅助气管插管滴注脂多糖）、IC组（气管暴露法滴注等渗NaCl溶液）、IL组（气管暴露法滴注脂多糖），每组各8只。记录制作动物操作时间。滴注脂多糖24 h后采集动脉测定动脉血氧分压（PaO₂）及氧合指数，计算肺组织湿/干重比值（W/D），检测支气管肺泡灌洗液（BALF）中蛋白含量，血清中肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6），并进行弥漫性肺泡损伤（DAD）评分。 结果EC组、IC组、EL组及IL组大鼠手术操作时间分别为（212 ± 24）、（296 ± 53）、（233 ± 44）、（321 ± 56）s，四组大鼠间比较，差异有统计学意义（F = 9.808，P < 0.001），且与IC组和IL组比较，EC组、EL组制作动物模型的操作时间均明显缩短（P均< 0.05）。四组大鼠间PaO₂、氧合指数、肺W/D、BALF蛋白含量、血清TNF-α、IL-6及DAD评分比较，差异均有统计学意义（F= 124.752、123.920、73.775、65.922、48.342、419.548、655.623，P均< 0.001）。进一步两两比较发现，与EC组及IC组比较，EL组及IL组大鼠的PaO₂ [（104 ± 10）、（105 ± 9）、（54 ± 3）、（53 ± 4）mmHg]、氧合指数[（498 ± 48）、（502 ± 43）、（261 ± 17）、（255 ± 21）mmHg]均显著降低，肺W/D [（4.14 ± 0.16）、（4.36 ± 0.18）、（5.53 ± 0.31）、（5.58 ± 0.29）]、BALF蛋白含量[（0.39 ± 0.07）、（0.34 ± 0.05）、（2.19 ± 0.13）、（2.15 ± 0.11）g/L]、血清TNF-α [（177 ± 38）、（186 ± 51）、（414 ± 61）、（440 ± 74）ng/L]、IL-6 [（104 ± 11）、（113 ± 28）、（584 ± 42）、（603 ± 56）ng/L]及DAD评分[（0.90 ± 0.29）、（0.82 ± 0.38）、（11.65 ± 0.89）、（12.23 ± 0.97）分]均显著升高（P均< 0.05）。 结论采用内窥镜下辅助气管插管滴注脂多糖能简单、有效地建立大鼠ARDS模型。

Study on rats with acute respiratory distress syndrome induced by intratracheal instillation of lipopolysaccharide in different ways

ObjectiveTo explore the rat model with acute respiratory distress syndrome (ARDS) induced by instillation of lipopolysaccharide (LPS) in endoscopic assisted endotracheal intubation and tracheal exposure. MethodsTotally 32 Sprague Dawley rats were randomly divided into 4 groups: the EC group (instillation of isosmotic NaCl solution in endoscopic assisted endotracheal intubation), EL group (instillation of LPS in endoscopic assisted endotracheal intubation), IC group (instillation of isosmotic NaCl solution in tracheal exposure), and IL group (instillation of LPS in tracheal exposure), 8 rats in each group. The operation time among groups was recorded. The levels of arterial oxygen partial pressure (PaO₂) and oxygenation index were detected on 24 hours after instillation of LPS. The wet/dry weight ratio (W/D) of pulmonary tissue, protein content of bronchoalveolar lavage fluid (BALF), levels of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) in serum, and diffuse alveolar damage (DAD) scores were estimated and compared. ResultsThe operation time in the EC group, IC group, EL group and IL group were (212 ± 24), (296 ± 53), (233 ± 44), (321 ± 56) s, respectively. The operation time among four groups showed significant difference (F = 9.808, P < 0.001), and the operation time was much shorter in the EC group and EL group than in the IC group and IL group (all P < 0.05). The levels of PaO₂, oxygenation index, pulmonary W/D ratio, protein content of BALF, TNF-α, IL-6 and DAD scores all showed significant differences among four groups (F = 124.752, 123.920, 73.775, 65.922, 48.342, 419.548, 655.623; all P < 0.001). Compared with the EC group and IC group, the levels of PaO₂ [(104 ± 10), (105 ± 9), (54 ± 3), (53 ± 4) mmHg] and oxygenation index [(498 ± 48), (502 ± 43), (261 ± 17), (255 ± 21) mmHg] were much lower, and the pulmonary W/D [(4.14 ± 0.16), (4.36 ± 0.18), (5.53 ± 0.31), (5.58 ± 0.29)], protein content of BALF [(0.39 ± 0.07), (0.34 ± 0.05), (2.19 ± 0.13), (2.15 ± 0.11) g/L], TNF-α [(177 ± 38), (186 ± 51), (414 ± 61), (440 ± 74) ng/L], IL-6 [(104 ± 11), (113 ± 28), (584 ± 42), (603 ± 56) ng/L] and DAD scores [(0.90 ± 0.29), (0.82 ± 0.38), (11.65 ± 0.89), (12.23 ± 0.97)] were much higher in the EL group and IL group (all P < 0.05). ConclusionIt is simple and effective to establish a ARDS rat model by instillation of LPS with endoscopic assisted endotracheal intubation.