盐酸酸雾吸入性肺损伤大鼠模型的建立与评价

Establishment and evaluation of rat models with inhalation lung injury caused by hydrochloric acid mist

【目的】建立盐酸酸雾吸入性肺损伤大鼠模型并对其进行评价。【方法】(1)改变致伤时间和氯化氢浓度将72只大鼠分为9组,即a:10 min,100 mg/L;b:10 min,200 mg/L;c:10 min,400 mg/L;d:20 min,100 mg/L;e:20 min,200 mg/L;f:20 min,400mg/L;g:30 min,100 mg/L;h:30 min,200 mg/L;i:30 min,400 mg/L。确定致伤条件。(2)56只大鼠随机分为空白对照组、吸氧对照组和损伤(按照第一部分确定的条件)后40 min、6 h、24 h、48 h、7 d取材实验组,检测各组大鼠动脉血气的变化,测定肺的干湿重比(W/D),观察大鼠的肺部大体变化及HE染色观察肺组织病理改变,检测肺组织匀浆液中超氧化物歧化酶(superoxide dismutase,SOD)、髓过氧化物酶(myeloperoxidase,MPO)和丙二醛(malondialdehyde,MDA)的变化,ELISA检测血清中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白介素-10(interleukin-10,IL-10)的变化。【结果】(1)a、b、d三组,大鼠损伤较轻,苏醒快,c、f、g、h、i五组,大鼠损伤较重,均有大鼠死亡。e组无大鼠死亡,均24~48 h内苏醒。(2)血气分析结果显示40 min时Pa O2急剧下降,24 h达到最低,随后缓慢上升。Pa CO2下降程度在损伤后40 min^24 h之间具有统计学意义。肺W/D呈渐进性增加,48 h达最大值。组织病理学显示盐酸酸雾造成的大鼠肺损伤主要为中性粒细胞增多的间质性肺炎。肺组织中的氧化指标检测显示随时间变化MPO、MDA逐渐增加;SOD的活性一直呈下降趋势。ELISA结果显示血清中TNF-α、IL-10逐渐增加。【结论】当造模条件为氯化氢浓度200 mg/L,致伤时间为20 min时,可以建立稳定的大鼠肺损伤模型,且可靠易于复制。

[ Objective ] To establish and evaluate rat models with inhalation lung injury caused by hydrochloric acid mist. [ Methods ] （1） Ac- cording to the injury time and the concentration of hydrochloric acid, 72 rats were divided into 9 groups： a, 10 min, 100 mg/L;b, lO rain, 200 mg/L; c, 10 min, 400 rag/L; d, 20 min, 100 mg/L; e, 20 min, 200 mg/L; f, 20 min, 400 mg/L; g, 30 min, 100 mg/L; h, 30 min, 200 mg/L; i,,30 min, 400 mg/L, The injury conditions were determined. （2） A total of 56 rats were randomly divided into blank control group, 02 inhalation control group and experimental groups （according to the conditions in part one） in which the rats were sacrificed after 40 min, 6 h, 24 h, 48 h and 7 d. The changes of arterial blood gas and the wet to dry weight （W/D） ratio of lung were detected. The general morphology of rat lung and HE-dyeing pathological changes of lung tissues were observed. The changes of superoxide dismutase （SOD）, myeloperoxidase （MPO） and malondialdehyde （MDA） in lung homogenate were determined. The levels of tumor necrosis factor-α （TNF-α） and interleukin-10 （IL-10） in serum were detected by ELISA. [ Results ] （1） In the groups of a, b and d, rats were injured slightly and woke up fast. However, rats in the groups of c, f, g, h and i were injured seriously and some died. Rats in group e were all alive and woke up within 24-48 h. （2） The results of blood gas analysis indicated that PaO2 decreased rapidly at 40 min, reached the bottom at 24 h and then went up slowly. The descending changes of PaCO2 were of statistical significance from 40 min to 24 h after the injury. The lung W/D ratio gradually increased and reached the maximum at 48 h. Histopathological changes showed that lung injury caused by hydrochloric acid mist in rats was mainly neutrophil-increased interstitial pneumonia. The activity of SOD tended to descend, while the levels of MPO and MDA in lung homogenate increased gradually. The results of ELISA showed that the serum levels of TNF-α and IL-10 increased gradually. [ Conclusion ] When the concentration of hydrochloric acid is 200 mg/L and the injury time is 20 min, the established rat models of lung injury are stable, reliable and replicable.