BALB/c小鼠哮喘模型焦虑及抑郁状态的实验检测

Detection of anxiety and depression status of BALB/c asthmatic mice model

目的 检测BALB/c小鼠哮喘模型的焦虑及抑郁状态.方法 20只小鼠按随机数字表法随机分为对照组和哮喘组各10只;哮喘组用鸡卵白蛋白加氢氧化铝致敏和激发,建立BALB/c小鼠哮喘模型,对照组用等量磷酸盐缓冲液代替.采用无创法检测不同浓度乙酰甲胆碱激发下2组小鼠的增强呼吸间歇（Penh）值来反映气道反应性.用高架十字迷宫法和强迫游泳法评估2组小鼠合并焦虑、抑郁程度.回收支气管肺泡灌洗液（BALF）进行细胞学分类.结果 哮喘组和对照组小鼠Penh值均随乙酰甲胆碱浓度增加而增加,哮喘组小鼠在生理盐水和乙酰甲胆碱浓度为0、5 g/L激发后Penh值分别为0.43±0.04、0.41 ±0.05和0.44_±0.04,与对照组（0.42±0.03、0.39 ±0.03、0.43±0.04）差异均无统计学意义（P =0.290、0.652、0.723）.在乙酰甲胆碱浓度为10、15、20 g/L时,哮喘组小鼠Penh值分别为0.57±0.03、0.85 ±0.04、1.57±0.08,均显著高于对照组的0.45±0.08、0.57 ±0.06、0.82±0.09（P =0.001、0.000、0.000）.哮喘组小鼠总进臂次数[开臂进入次数（OE）＋闭臂进入次数（CE）]、OE占OE＋ CE百分比（OE％）、开臂滞留时间（OT）占总滞留时间[开臂滞留时间（OT）＋闭臂滞留时间（CT）]百分比（OT％）分别为（18.3±3.6）次、（22.2±3.1）％、（16.7±4.2）％,显著低于与对照组的（24.0±2.9）次、（28.0±3.4）％、（21.8±4.6）％（P=0.001、0.001、0.019）;游泳不动时间为（147±12）s,显著高于对照组的（133±10）s （P =0.010）;BALF中细胞总数及嗜酸粒细胞比值[（10.0±4.0）×105/ml和（68.18±3.76）％]均显著高于对照组[（1.7±0.4）×105/ml和（0.12±0.07）％]（均P=0.000）.结论 BALB/c哮喘小鼠合并焦虑、抑郁状态,可应用该模型研究哮喘与焦虑、抑郁共病现象的分子生物学机制.

Objective To detect the anxiety and depression status of BALB/c asthmatic model in mice.Methods A total of 20 mice were randomly divided into asthma and control groups （n =10 each）.The animals were sensitized and challenged with ovalbumin for asthmatic model.Phosphate buffer solution was used in the control group.The mice were challenged with increasing concentrations of methacholine aerosol and the airway resistance was measuring non-invasively by enhanced pause （Penh）.And elevated plus maze test system and forced swim were employed to evaluate their anxiety and depression status.Then bronchoalveolar lavage cytology was detected for evaluating the airway inflammation.Results In both groups,Penh value increased with the rising concentration of methacholine.The Penh values of asthmatic mice exciting with normal saline and methacholine concentration of 0,5 g/L were 0.43 _± 0.04,0.41 _± 0.05 and 0.44 ± 0.04 versus 0.42 ± 0.03,0.39 ± 0.03,0.43 _± 0.04 in control group.The difference had no statistical significance （P =0.290,0.652,0.723）.At the concentration of methacholine 10,15,20 g/L,the Penh values of asthmatic mice were 0.57 ± 0.03,0.85 ± 0.04,1.57 ± 0.08 and they were significantly higher than those of control group （0.45 ± 0.08,0.57 ± 0.06,0.82 ± 0.09） （P =0.001,0.000,0.000）.Asthmatic mice in the elevated plus maze on a behavioral test and found that asthmatic mice open arm entries sum closed arm entries （OE ＋ CE）,percent of open arm/total entries （OE%） and percent of open arm/total time （OT%） was （18.3 ± 3.6） times,（22.2 ± 3.1） %,（16.7 ±_ 4.2） %.They were significantly lower than those of control group （（24.0 ± 2.9） times,（28.0 ± 3.4）%,（21.8 ± 4.6）%） （P =0.001,0.001,0.019）.Asthmatic mice swimming time was significantly higher than that of control group （147 ± 12） vs （133 ±_ 10） s （P =0.010）.The total number of cells and eosinophil ratio in asthma group was （10.0 ±4.0） × 105/ml and （68.18 _± 3.76）%.And they were significantly larger than those of control group （（1.7 __ 0.4） × 105/ml,（0.12 ± 0.07） %） （both P =0.000）.Conclusion Asthmatic mice with concurrent anxiety and depression may be a model for elucidating the molecular mechanisms of asthma with anxiety and depression.