噻托溴铵对慢性阻塞性肺疾病大鼠膈肌细胞凋亡的影响

The influence of tiotropium bromide on the apoptosis of diaphragmatic muscle in chronic obstructive pulmonary disease rats

目的探讨噻托溴铵对COPD大鼠膈肌细胞过度凋亡的调节作用及其机制。方法60只雄性Wistar大鼠随机分成健康对照组、COPD组和噻托溴铵干预组。采用烟熏及气管内注射脂多糖建立COPD模型，第91天将大鼠处死，观察各组大鼠肺组织的病理变化，TUNEL法检测各组膈肌细胞的凋亡率，免疫组织化学法检测膈肌细胞内Fas蛋白的表达，Westrenblot法检测膈肌细胞中FasL及Caspase-3的含量，RT—PCR法检测Caspase-3基因表达。结果噻托溴铵组大鼠膈肌细胞的凋亡率、Fas、FasL、Caspase-3蛋白含量及Caspase-3基因表达均低于COPD组，高于健康对照组，3组之间差异有统计学意义（Pd0．05）。结论cOPD的发生发展过程中存在膈肌细胞过度凋亡，Fas／FasL途径参与了膈肌细胞凋亡的调控，噻托溴铵可抑制膈肌细胞的过度凋亡，延缓COPD的进程。

Objective Explore the influence and mechanism of tiotropium bromide on the excessive apoptosis of diaphragmatic muscle in chronic obstructive pu].monary disease （COPD） rats. Methods Sixty male Wistar rats are randomly divided into control group, COPD group and tiotropium Bromide group. The rat models of COPD were established by passive smoking as well as intratracheal instillation of lipopolysaccharide respectively. The pathologic change of the lung tissue and airway were observed by HE staining. The diaphragmatic muscle cell apoptosis rate, Fas, FasL and Caspase 3 protein and Caspase-3 gene expression of diaphragmatic muscle cell were measured. Results HE staining proved that the rat COPD models were successfully established. The COPD group appears obvious emphysema, while the riotropium bromide group appears mild emphysema. The apoptosis rate, Fas, FasL and Caspase-3 protein and Caspase-3 gene expression of diaphragmatic muscle cell in tiotropium bromide group were higher than the healthy control group, but less than COPD group （ P〈0.05）. Conclusions Excessive apoptosis of diaphragmatic muscle cell was present in rats with COPD. Fas/FasL mediated apoptosis way was involved in diaphragm apoptosis. Tiotropium bromide can reduce the level of apoptosis in diaphragmatic muscle cell.