李斌愷 賴克方 洪燕華 王法霞 陳如沖 林少建 鐘南山
【摘要】 目的 目前國內對支氣管哮喘 (簡稱哮喘) 小鼠的評價多數僅立足于氣道炎性指標 ,不能完全反映哮喘的病理生理特征 。 本所率先從國外引進了小動物無創檢測和有創檢測肺功能儀 。 無創法檢測時小鼠不必麻醉 ,而且每次可以同時檢測多只小鼠 ,具有較大的優越性 ,但能否取代有創法尚需更多的數據 。本研究旨在建立無創檢測小鼠氣道高反應性的檢測方法 ,并與有創檢測方法進行比較 。 方法 根據動物模型和氣道反應性檢測方法不同 ,動物分為 : ① 無創哮喘組 ; ② 無創對照組 ; ③ 有創哮喘組 ; ④ 有創對照組 。 采用卵白蛋白致敏和激發 ,建立 BALB/ c 小鼠哮喘模型 ,生理鹽水作為對照 ,分別用無創和有創的方法測定氣道反應性 。 哮喘動物霧化吸入0 . 2～50 g/ L 倍增濃度的乙酰甲膽堿 ( Mch) ,測定相應濃度下的增強呼氣間歇 ( Pe nh) 值或氣道阻力 ( RL ) 值等指標 。 將小鼠吸入 Mc h 后 RL 或 Penh 增加 2 倍的激發濃度以 PC100 來表示 。 所有動物都行支氣管肺泡灌洗 , 收集灌洗液 , 涂片染色后分類計數 。結果 無創哮喘組 PC100 均 ≤6 . 25 g/ L ,對照組 PC100 均 ≥ 12 . 5 g/ L 。 其 Lo g2 ( 10 PC100 ) 值 ( 5 . 36 ± 0 . 84) 顯著低于對照組 ( 7 . 97 ± 0 . 82)( P < 0 . 01) 。 無創哮喘組從 Mc h 濃度3 . 12 g/ L 開始 , 其 Pe nh 值明顯高于對照組 。有創哮喘組 RL 值從Mc h 濃度0 . 39 g/ L開始就明顯高于相應對照組 ( P < 0 . 05) 。 無創組與有創組的氣道反應性相關系數 R =0 . 96 ( P < 0 . 01) 。 無創哮喘組和有創哮喘組的嗜酸粒細胞分別為 ( 54 . 00 ± 5 . 96) % , ( 55 . 93 ± 5 . 92) % ,顯著高于各自對照組的 ( 0 . 38 ± 0 . 52) % , ( 0 . 63 ± 0 . 74) %( P < 0 . 01) 。 結論 本研究表明以 Penh 為主要測定指標的無創方法 ,可以成功檢測哮喘小鼠氣道高反應性 。
【關鍵詞】 小鼠 ;氣道反應性 ; 支氣管哮喘 ;增強呼氣間歇 ; 無創檢測
【Ab st ra ct】 Objective Most of t he evaluation on asthmatic mouse model only based on the airway inflammation , not fully reflecting its pathophysiology. Noninvasive method of measuring bronchial hyper responsiveness can detect several conscious mice in one time , but whether it can replace the invasive technique needs more data to be identified. This study aims to establish a non-invasive detection of mouse airway hyper responsiveness ,comparing with the invasive method. Methods According to different ways of detection , mice were divided into four groups : non-invasive asthma group , non-invasive control group ,invasive asthma group and invasive control group . Mouse was sensitized and challenged with ovalbumin ( OVA) for asthmatic model . The mice were challenged with increasing concentrations of methacholine aero sol from 0 . 2 250 g/ L ,and the airway resistance was measured. Enhance pause ( Penh) or airway resistance ( RL ) was taken for each group . The dose of methacholine causing 100 % increase of respiratory resistance was defined as PC100 . The PC100 values were converted into Log2 ( 10 PC100 ) for statistical analysis. Bro nchoalveolar lavage cytology was performed to evaluate the airway inflammation. Results The PC100 of non-invasive asthma group was ≤6 . 25 g/ L , and the non-invasive control group’s ≥12 . 5 g/ L . I t s Lo g2 ( 10 PC100 ) value was significantly less than that of control group , ( 5 . 36 ± 0 . 84) vs ( 7 . 97 ± 0 . 82) ( P < 0 . 01) . The Penh value of non -invasive asthma group began significantly increasing when the concentration of Mch was 3 . 12 g/ L . The RL value of invasive asthma group was increasing at the beginning of 0 . 39 g/ L ( P < 0 . 05) . The coefficient rate of two groups was 0 . 96 ( P < 0 . 01 ) . The EOS ( %) of non-invasive asthma group or invasive asthma group was 54 . 00 ± 5 . 96 , 55 . 93 ± 5 . 92 respectively , significantly larger than that of control group ( 0 . 38 ±0 . 52 ,0 . 63 ± 0 . 74 , P < 0 . 01) . Conclusions This study shows that single-chambered barometric whole-body plethysmography as a non- invasive lung function test can success fully detect airway responsiveness in asthmatic mouse model .