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摘要:
背景 纤毛在气道防御机制中发挥着重要作用。由于观察技术和设备的限制，既往关于鼻用糖皮质激素对纤毛运动影响的研究主要是基于离体试验，尚无相关的在体动物研究。目的：使用一种新的方法观察并比较鼻用糖皮质激素对在体和离体鼻黏膜纤毛运动的影响。方法：60只大鼠随机分为在体观察和离体观察2组，每组又随机分为布地奈德、糠酸莫米松和丙酸氟替卡松3个亚组，各亚组均以磷酸盐缓冲液(phosphate buffered saline, PBS)作为对照，每个试验组和对照组各5只大鼠。在体观察组将大鼠麻醉后立即手术暴露鼻中隔黏膜；离体观察组将大鼠过量麻醉后断头处死，取其鼻中隔黏膜放置于培养基中。两组皆放置于新型数码显微系统下观察鼻黏膜表面的纤毛运动情况，计算并比较60 min内在体和离体观察组大鼠鼻黏膜表面滴加药物前后纤毛摆动频率(ciliary beat frequency, CBF)的差异。结果：在体观察组大鼠鼻黏膜的CBF显著高于离体观察组大鼠鼻黏膜的CBF[(9.83 ± 0.94)Hz vs (6.10 ± 0.91)Hz, P＜0.001)]。在体观察发现，布地奈德、糠酸莫米松和丙酸氟替卡松在60 min内对大鼠鼻黏膜CBF未见明显的影响。而离体观察条件下，布地奈德使CBF即刻下降，在60min内组间各时间点的CBF差异均有统计学意义(P＜0.05)；糠酸莫米松使CBF持续下降，且加药5 min后组间各时间点的CBF差异均有统计学意义(P＜0.05)；丙酸氟替卡松使CBF波动下降，且在0 ~ 5 min和20 ~ 60 min内组间各时间点的CBF差异均有统计学意义(P＜0.05)。结论：布地奈德、糠酸莫米松 [1] 和丙酸氟替卡松对在体鼻黏膜的纤毛运动无显著的影响，而对离体鼻黏膜的纤毛运动均产生了不同程度的抑制作用，这表明鼻用糖皮质激素无明显的纤毛毒性，具有良好的安全性；同时本研究所使用的在体评估系统弥补了离体观察与机体真实环境差异较大的不足，可用于客观评价鼻用药物的安全性及纤毛毒性。Abstract:Background Cilia plays an important role in airway defense mechanisms. However, due to limitations in the research techniques and tools for observation, in vivo animal studies on the effects of intranasal corticosteroids on ciliary motion are scarce. These researches have mainly been performed in vitro. Objective To investigate the effects of intranasal corticosteroids on nasal ciliary motion in vivo and in vitro using a new observation method. Methods Sixty rats were randomly divided into in vivo observation group and in vitro observation group. Each group was randomly divided into three subgroups: budesonide, mometasone furoate and fluticasone propionate, which was treated with phosphate buffered saline (PBS) as control. Rats were anesthetized and immediately surgically exposed the nasal septal mucosa in vivo observation group; while rats were over-anesthetized, severed and executed, and their nasal septal mucosa was taken and placed in the culture medium in vitro observation group. Both groups were placed under the new digital microscope system to observe the ciliary motion on the nasal mucosal surface. The differences in ciliary beat frequency (CBF) between the two groups were compared before and after drug addition to the nasal mucosal surface of rats within 60 min. Results The CBF of nasal mucosa of the rats in vivo observation group was significantly higher than that in vitro observation group ([9.83 ± 0.94]Hz vs [6.10 ± 0.91]Hz, P<0.001). Under in vivo conditions, budesonide, mometasone furoate, and fluticasone propionate had no significant effect on CBF in rat nasal mucosa within 60 min. Under in vitro conditions, budesonide caused an immediate decrease in CBF, with statistically significant differences in CBF between two groups at all time points within 60 min (P<0.05); mometasone furoate decreased CBF continuously, and the difference in CBF between two groups at each time point after 5 min of addition was statistically significant (P<0.05); fluticasone propionate caused a fluctuating decrease in CBF, and the difference in CBF between two groups at all time points from 0 to 5 min and from 20 to 60 min was statistically significant (P<0.05). Conclusion Budesonide, mometasone furoate, and fluticasone propionate have no significant effect on ciliary motion in vivo, while they all produce varying degrees of inhibition of ciliary motion in vitro, indicating that intranasal corticosteroids have no significant ciliary toxicity and good safety. Meanwhile, the in vivo evaluation system used in this study compensates for the great difference between in vitro observation and the real physiological environment and can be used to objectively evaluate the safety and ciliary toxicity of nasal drugs. -
Key words:
- intranasal corticosteroids /
- ciliary beat frequency /
- nasal mucosa /
- rats /
- digital microscope system
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图 1 在体和离体观察鼻黏膜的CBF比较(n=30)
Figure 1. Comparison of CBF of nasal mucosa observed in vivo and in vitro (n=30)
图 2 布地奈德和PBS对在体和离体鼻黏膜CBF的影响
A:布地奈德对在体CBF的影响；B:布地奈德对离体CBF的影响(n=5, bP＜0.01, vs PBS)
Figure 2. Effect of budesonide and PBS on CBF of nasal mucosa in vivo and in vitro
A: Effect of budesonide on in vivo CBF; B: Effect of budesonide on in vitro CBF (n=5, bP＜0.01, vs PBS)
图 3 糠酸莫米松和PBS对在体和离体鼻黏膜CBF的影响
A: 糠酸莫米松对在体CBF的影响；B: 糠酸莫米松对离体CBF的影响(n=5, aP＜0.05, bP＜0.01, vs PBS)
Figure 3. Effect of mometasone furoate and PBS on CBF of nasal mucosa in vivo and in vitro
A: Effect of mometasone furoate on in vivo CBF; B: Effect of mometasone furoate on in vitro CBF (n=5, aP＜0.05, bP＜0.01, vs PBS)
图 4 丙酸氟替卡松和PBS对在体和离体鼻黏膜CBF的影响
A: 丙酸氟替卡松对在体CBF的影响；B: 丙酸氟替卡松对离体CBF的影响(n=5, aP＜0.05, bP＜0.01, vs PBS)
Figure 4. Effect of fluticasone propionate and PBS on CBF of nasal mucosa in vivo and in vitro
A: Effect of fluticasone propionate on in vivo CBF; B: Effect of fluticasone propionate on in vitro CBF (n=5, aP＜0.05, bP＜0.01, vs PBS)
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