氨溴索对吸烟大鼠支气管黏膜超微结构及BALF中IL-8 TNF-α的影响

Effect of ambroxol on the bronchil mucosa ultrastructure and IL-8 TNF-α in BALF in smoking rats

目的:观察氨溴索对被动吸烟大鼠气道炎症的作用。方法:实验大鼠随机分为:健康对照组、吸烟模型组和氨溴索干预组。采用吸香烟法建立大鼠支气管炎模型。氨溴索干预组自模型开始建立时给予氨溴索片40mg-1.kg-1.d-1,分2次喂食。2个月后,扫描电镜观察各组气道的超微结特点,测定支气管肺泡灌洗液(BALF)中白细胞介素-8(IL-8)和肿瘤坏死因子α(TNF-α)的水平,瑞氏染色进行支气管肺泡灌洗液细胞计数和分类。结果:模型组大鼠的支气管上皮细胞纤毛明显脱失、减少、紊乱、倒伏、扭曲及粘连,杯状细胞表面微绒毛减少,上皮细胞萎缩,断裂甚或脱失,炎性细胞浸润。氨溴索干预组与健康对照组相比,气管、支气管上皮细胞超微结构仍具有病理改变。但镜下改变较吸烟模型组明显减轻;吸烟模型组与氨溴索干预组BALF中细胞总数、中性粒细胞数及百分比与健康对照组相比明显增高(P<0.01),但吸烟模型组与氨溴索干预组细胞数差异无显著性;吸烟模型组与健康对照组比较BALF中IL-8和TNF-α的水平显著增高(P<0.01),氨溴索干预组BALF中IL-8和TNF-α的水平与吸烟模型组比较降低(P<0.01)。结论:吸烟引起大鼠支气管黏膜上皮细胞纤毛明显脱失,杯状细胞表面微绒毛减少,炎性细胞浸润,BALF中IL-8、TNF-α、细胞计数增多,应用氨溴索的大鼠气道炎症减轻,氨溴索对吸烟引起的大鼠气道支气管炎有明显的保护作用。

Objective：To observe the effect of ambroxol on chronic bronchitis in smoking model of rat. The changes of ultrastructures of bronchus, cell count and the level of interleukin-8 （IL-8） and tumor necrosis factor-a （TNF-α） in the bronchoalveolar lavage fluid （BALF） in animal were observed. Method： Rats were randomly divided into three groups： 1 .control group （ n = 10） ; 2. smoking group （ n = 10） ; 3. Ambroxol intervention group（ n = 10）. Chronic bronchitis in group （2） and （3） established by passive cigarette smoking. In group 0, ambroxol was administrated 40 mg/kg daily for 2 months before the rats were dead. The ultrastructures of bronchial mucosa were observed by scanning electron microscope. The levels of IL-8 and TNF-α and cell count in BALF was detected by ELISA and Wright＇ s staining separately. Result：Smoking group manifested epithelial cell cilia of bronchial mucosa lossing, lodging and twisting, microvilli of chalice cell decreasing, inflammatory cell infiltrating, epithelial cell atrophing, chalice cell in region increasing , microvillus decresing. The surface of ciliated cell is thin and fiat. These feature correspond with the of human chronic bronchitis. Compared with the control group , the ambroxol intervention group appeared pathological changes of tracheal and bronchial epithelium cell. The numbers of total cells and neutrophil in BALF of the smoking group and the ambroxol group were significantly increased compared to the control group （P 〈 0.01 ）. The numbers of total cells and neutrophil in BALF of the ambroxol group were significantly decreased compared to the smoking group （P 〈 0.05）. The level of IL-8 and TNF-α in BALF of the smoking group was significantly increased compared to the control group（ P 〈 0.01）. whereas the ambroxol group was significantly decreased compared to the smoking group （P 〈 0.01 ）. Conclusion：Smoking could cause obviously bronchial mucosa to loss epithelial cell cilia, decrease microvilli chalice cell, increase of IL-8, TNF-α, cell counting in BALF. Inflammation of airway could be alleviated by using ambroxol.