改良式小鼠颈透视下插管滴定脂多糖制备急性肺损伤模型的研究

Study on the use of improved endotracheal intubation to instill lipopolysaccharide for the prepara- tion of mouse acute lung injury model

目的探索一种损伤小、可靠、高效的小鼠气管内插管方法并滴定脂多糖制备急性肺损伤模型。方法80只BALB／C小鼠按随机数字表法分为两组：脂多糖组（LPS组）和对照组（Ns组）。气管内插管成功后分别滴定脂多糖（3mg／kg）和生理盐水（1．5ml／kg），记录插管一次成功率、最终成功率、存活率。24h后计数左肺肺泡灌洗液（BALF）中总细胞数，瑞氏染色进行细胞分类计数，BCA法测定总蛋白浓度，肺组织烘干后计算湿／于比，动脉血气分析测定血氧分压（PaO2）并计算氧合指数。结果改良式颈透视下插管一次成功率92．5％、最终成功率和存活率均为100％；LPS组BALF中总细胞数[（10．82±3．51）×105／mlVS（0．72±0．52）×105／ml，t=-6．294，P〈0．01]、多形核粒细胞占总细胞数比例[（93．93±1．77）％VS（2．2±0．91）％，t=-105．565，P〈0．01]、单个核粒细胞占总细胞数比例[（6．07±1．77）％vs（97．8±0．91）％，t=-105．565，P〈0．01]、总蛋白浓度[（0．49±0．13）mg／mlVS（0．29±0．11）mg／ml，t=-2．823，P〈0．05]、湿／干比值（4．60±0．18VS4．16±0．25，t=-4．793，P〈0．01）、动脉PaO2【（68．57±7．23）％VS（87．00±6．33）％，t=4．571，P〈0．01]和氧合指数[（326．53±34．43）mmHgVS（414．29±30．16）mmHg，t=4．571，P〈0．01]与NS组比较差异均有统计学意义。结论改良式小鼠颈透视下气管内插管成功率高、损伤小，且滴定LPS3mg／kg成功制备了急性肺损伤模型。

Objective To explore a minimally invasive, reliable, and efficient method for endotra- cheal intubation to instill lipopolysaccharide （LPS） for preparation of acute lung injury （ALI） in mice. Methods A total of 80 BALB/C mice was randomly selected into LPS group （ n = 40） and control group （Normal saline, NS; n = 40）. After a successfully endotracheal intubation, each mouse was instilled by LPS （3 mg/kg） in LPS group, and NS （1.5 ml/kg） in NS group, respectively. The one-time success rate and final success rate of the endotracheal intubation, and survival rate were recorded. After 24 hours, the total number of cells in bronehoalveolar lavage fluid （BALF） of left lung was counted with light microscope. The cells were classified and counted after Wrightg stain. Total protein concentration in BALF was assayed with a BCA kit. Wet/dry value was calculated after the lung became dry. Artery blood PaO2 was tested and the oxygenation index was counted. Results Compared to NS group, the LPS group had the one-time success rate 92. 5% , and the final success rate 100%, survival rate 100%, the total number of ceils [ （ 10. 82 ± 3.51 ） ×105/ml vs （0. 72 ＋ 0. 52） × 105/ml, t = - 6. 294, P 〈 0. 01 ] , the rate of polymorphonulear leukocytes in total cells[ （93.93 ±1.77）% vs （2.2 ＋0.91）%, t = -105.565, P 〈0.01], the rate of mononuclear leukocytes in total cells[ （6. 07 ± 1.77） % vs （97. 8±0. 91 ） % , t = - 105. 565, P 〈0. 01 ] , total protein concentration [ （0.49 ±0. 13 ） mg/ml vs （0. 29 ＋ 0. 11 ） mg/ml, t = - 2. 823, P 〈 0. 05 ], W/D ratio（4. 60 ±0. 18 vs 4. 16 ±0. 25, t = -4. 793, P 〈0. 01）, PaO2[ （68. 57 ±7. 23）% vs（87.00± 6. 33）%, t =4. 571, P 〈0. 01 ], and oxygenation index[ （326. 53 ± 34. 43） mmHg vs（414. 29 ±30. 16） mmHg, t = 4. 571, P 〈 0. 01 ]. Conclusions Improved method for endotracheal intubation has high success rate and minimal injury, and instillation of LPS （3 mg/kg）can induce mice ALI successfully.