小鼠鼻腔及气管黏膜上皮细胞的构成和纤毛反应性比较研究

A Comparative Study on Mouse Nasal and Tracheal Epithelium for Cellular Composition and Ciliary Cell Reactivity

目的比较小鼠鼻腔及气管黏膜上皮细胞的构成以及纤毛摆动频率对外源性刺激的反应性。方法 采用酶消化法获取小鼠鼻腔及气管黏膜纤毛细胞,观察纤毛细胞的得率以及ATP和苯扎溴铵对纤毛摆动频率的影响;采用免疫组化染色及扫描电镜观察鼻腔及气管黏膜上皮的细胞构成。结果 ①培养细胞中小鼠鼻甲的纤毛细胞得率可达(45±5)%,气管组织仅有(15±5)%;②免疫组化染色鼻腔及气管黏膜上皮细胞均有乙酰化α-tubulin阳性表达,鼻腔的纤毛细胞层更连续,细胞更密集;③100μmol/LATP均能引起鼻甲及气管的纤毛摆动频率增加,增加率分别为235.06%和118.49%;0.005%苯扎溴铵均能使2个部位的纤毛摆动频率下降,降至原有频率的50.31%和8.27%。2组频率比较差异均有统计学意义,P值分别为0.013和0.004;④扫描电镜观察,鼻腔黏膜上皮中纤毛细胞的比例(90%)明显高于气管上皮纤毛细胞比例(30%～40%)。结论 小鼠鼻腔和气管黏膜上皮中纤毛细胞的比例存在明显差异;2个部位的纤毛摆动频率对外源性刺激的反应程度存在明显差异;从纤毛细胞的数量上看,鼻腔远远高于气管,其中鼻甲是个不容忽视的丰富资源。

Objective：To compare the proportion of cell populations and effects of exogenous stimuli on ciliary beat frequency（CBF） in mouse nasal and tracheal epithelium.Methods：Mouse nasal and tracheal epithelial cells were cultured by enzymatic digestion culture method,the yield of ciliary cells and also the effects of ATP,benzalkonium bromide on CBF were observed.In addition,the proportion of cell populations in nasal and tracheal epithelium was investigated by immunohistochemistry and scanning electron microscope（SEM）.Results：① Cultured ciliated epithelial cells were（45±5）% of the total cell population in nasal turbinate epithelium,whereas,tracheal ciliary cells comprised（15±5）% of total cell population.② Expression of acytylated α-tubulin was detected in both nasal and tracheal epithelium,however,the positive cells in nasal epithelium appeared more continuously and densely than in tracheal epithelium.③ CBF increased with treatment of 100 μmol/L ATP both in nasal（235.06%） and tracheal（118.49%） epithelial cells,the difference between two groups was significant（P=0.013）.While CBF decreased with addition of 0.005% benzalkonium bromide,significant difference between nasal（50.31%） and tracheal（8.27%） epithelium was detected（P=0.004）.④ By SEM,the proportion of ciliary cells in nasal（90%） epithelium was much higher than in trachea（30%~40%）.Conclusion：There are significant differences in the proportion of ciliary cells and CBF changes towards exogenous stimuli in nasal and tracheal epithelium;ciliary cell numbers of nasal epithelium were much higher than those in trachea,among which turbinate is an abundant resource for ciliary cell cultures that should not be ignored.