论文部分内容阅读
目的观察布地奈德(BUD)、卡介苗(BCG)干预对支气管哮喘小鼠Ⅱ型肺泡上皮细胞(ATⅡ)损伤的影响。方法昆明小鼠40只随机分为哮喘模型组、BUD干预组、BCG干预组和正常对照组。以卵白蛋白致敏和激发复制小鼠哮喘模型,并行BUD、BCG干预。HE染色观察其呼吸道周围嗜酸性粒细胞(EOS)浸润。ELISA法检测其血清、支气管肺泡灌洗液(BALF)中Krebs vonden lungen-6(KL-6)表达及BALF中IL-4、IFN-γ表达。免疫组织化学染色(SABC法)观察其肺表面活性蛋白(SP-A、SP-B)的表达。透射电镜观察其ATⅡ形态。结果哮喘模型组、BUD干预组、BCG干预组、正常对照组EOS分别为(806±91)个.mm-2、(173±25)个.mm-2、(189±19)个.mm-2及(3±1)个.mm-2;血清KL-6水平分别为(1.514±0.027)ng.L-1、(1.101±0.020)ng.L-1、(1.008±0.019)ng.L-1及(0.753±0.033)ng.L-1;BALF中KL-6水平分别为(0.842±0.020)ng.L-1、(0.268±0.024)ng.L-1、(0.243±0.026)ng.L-1及(0.090±0.015)ng.L-1;BALF IL-4水平分别为(22.61±2.35)ng.L-1、(13.34±2.41)ng.L-1、(15.56±3.08)ng.L-1及(0.62±0.15)ng.L-1;BALF IFN-γ水平分别为(40.56±3.58)ng.L-1、(60.37±6.41)ng.L-1、(57.87±6.28)ng.L-1及(75.64±6.09)ng.L-1;SP-A IOD值分别为45.89±10.08、181.34±29.41、200.12±32.08及389.62±45.05;SP-B IOD值分别为10.34±2.42、88.76±12.66、87.13±14.58及187.89±27.15。上述指标除BUD干预组与BCG干预组之间差异无统计学意义外,余各组间差异均有统计学意义(Pa<0.01)。电镜下可见哮喘模型组小鼠ATⅡ板层小体和线粒体减少,有明显排空和空泡化改变,部分细胞变性、坏死。干预后上述改变均有明显改善。哮喘模型组、BUD干预组及BCG干预组BALF中KL-6水平与EOS呈明显正相关,与BALF中IL-4/IFN-γ亦呈明显正相关。SP-A与BALF IL-4/IFN-γ呈明显负相关。结论支气管哮喘使小鼠ATⅡ明显受损,KL-6表达增加,SP-A、SP-B表达下降。BUD、BCG干预可减轻哮喘ATⅡ的损伤。
Objective To observe the effect of BUD and BCG on the injury of type Ⅱ alveolar epithelial cells (AT Ⅱ) in bronchial asthma mice. Methods Forty Kunming mice were randomly divided into asthma model group, BUD intervention group, BCG intervention group and normal control group. To ovalbumin sensitized and stimulated replication of mouse asthma model, parallel BUD, BCG intervention. HE staining was used to observe the infiltration of eosinophils (EOS) around the airway. ELISA was used to detect the expression of KL-6 and IL-4, IFN-γ in BALF in serum and bronchoalveolar lavage fluid (BALF). Immunohistochemical staining (SABC method) was used to observe the expression of pulmonary surfactant protein (SP-A, SP-B). Transmission electron microscopy AT Ⅱ morphology. Results The EOS of asthmatic model group, BUD intervention group, BCG intervention group and normal control group were (806 ± 91) mm-2, (173 ± 25) mm-2 and (189 ± 19) mm- (1.514 ± 0.027) ng.L-1, (1.101 ± 0.020) ng.L-1, (1.008 ± 0.019) ng.L (-1) and (0.753 ± 0.033) ng.L-1 respectively. The levels of KL-6 in BALF were (0.842 ± 0.020) ng.L-1, (0.268 ± 0.024) ng.L- . The levels of IL-4 in BALF were (22.61 ± 2.35) ng.L-1, (13.34 ± 2.41) ng.L-1 and (15.56 ± 3.08) (40.56 ± 3.58) ng.L-1, (60.37 ± 6.41) ng.L-1, (57.87 ± 6.28) ng.L-1 and (0.62 ± 0.15) ng.L- ) and the SP-A IOD values were 45.89 ± 10.08, 181.34 ± 29.41, 200.12 ± 32.08 and 389.62 ± 45.05, respectively; the IOD values of SP-B were 10.34 ± 2.42, 88.76 ± 12.66, 87.13 ± 14.58 and 187.89 ± 27.15. Except for the BUD intervention group and the BCG intervention group, there was no significant difference between these two groups (Pa <0.01). Electron microscopy showed that the asthma model group ATⅡ lamellar body and mitochondria decreased significantly evacuated and vacuolar changes, some cells degeneration and necrosis. After the intervention, these changes have been significantly improved. The level of KL-6 in BALF was positively correlated with EOS in asthmatic model group, BUD intervention group and BCG intervention group, and positively correlated with IL-4 / IFN-γ in BALF. SP-A and BALF IL-4 / IFN-γ showed a significant negative correlation. Conclusions Bronchial asthma causes obvious damage to AT Ⅱ, increases the expression of KL-6 and decreases the expression of SP-A and SP-B. BUD, BCG intervention can reduce AT Ⅱ injury in asthma.