目的探讨Ⅱ型肺泡上皮细胞(AECⅡ)凋亡在被动吸烟肺气肿大鼠肺功能下降中的相关作用。方法选取907,清洁级雄性sD大鼠,完全随机平均分成6组,A组正常对照组,B、C、D、E、F组大鼠被动吸入不同浓度香烟烟雾。采用小动物肺功能仪测定各组大鼠肺功能、肺组织病理学检测、Real-timePCR测定肺泡表面活性蛋白A(SPA)和SPCmRNA水平、免疫组织化学法检测SPA和SPC蛋白表达水平,TUNEL与SPC双重免疫荧光染色分析AECⅡ凋亡情况。结果 当香烟烟雾中一氧化碳(CO)浓度达到(300±10)ppm以上时,大鼠肺功能中气道阻力(AR)明显增加、肺动态顺应性(Cdyn)及呼气峰值流速(PEF)降低(P〈0.05);大鼠肺组织出现明显肺气肿样表现,肺泡平均内衬间隔(MLI)和平均肺泡面积(MAA)显著增加(P〈0.05);肺组织中SPA和SPCmRNA及蛋白表达水平明显下降(P〈0.05),AECⅡ凋亡百分比显著增高(P〈0.05)。结论AECII凋亡在被动吸烟建立的大鼠肺气肿发生发展过程中起重要作用。
Objective To study the effects of type II alveolar epithelial cells apoptosis on pulmonary function decline in rats with emphysema. Methods Ninety SPF-grade male Sprague-Dawley rats were randomly sorted into six groups. A group was control group; rats form B, C, D, E and F groups were exposed to different passive smoking concentrations. Small animal lung function instrument was used to analyze the pulmonary function of each rat. Histological changes of rats were analyzed by H&E staining, and surfactant protein A ( SPA ) and SPC mRNA expressions were detected by Real-time PCR; SPA and SPC protein expressions were measured by immunohistochemical analysis. TUNEL/SPC immunofluorescence staining was used to analyze the apoptosis of type II alveolar epithelial cells ( AEC Ⅱ ) . Results When the concentration of carbon monoxide ( CO ) in cigarette smoke was above 300 + 10 ppm, the airway resistance ( AR ) from rats was increased significantly, and the pulmonary dynamic compliance and peak expiratory flow were obviously decreased in lung function ( P〈0.05 ) . Enlargement of airspace was observed and the amount of alveoli was decreased in the lung tissues, and the mean linear intercept and mean alveolar airspace from rats were significantly increased ( P〈0.05 ) ; SPA and SPC mRNA and protein expressions were markedly decreased ( P〈0.05 ) ; and the percentage of AEC Ⅱ apoptosis was significantly increased ( P〈0.05 ) . Conclusions AEC Ⅱ apoptosis plays an important role in the development of emphysema in rats induced by passive smoking.