In vitro effect of combined hypertonic saline and salbutamol on ciliary beat frequency of human nasal epithelial cells

Abstract Acute infectious diseases of the respiratory tract are regularly associated with increased concentrations of neopterin in serum and/or urine. Most likely, the pteridine compound is not only a marker of an activated cellular immune system but may exert distinct biochemical functions in the process of infections as well. In response to viral contact, macrophages located in the nasal mucosa may represent a source for neopterin in the nasal cavity with potential hazardous effects on ciliary beat frequency (CBF), thus promoting chronic infectious processes. The aim of the present study was to investigate the effects of neopterin at different concentrations (10 nM, 100 nM, and 100 μM) on CBF in human nasal epithelial cells in vitro. Samples were taken from the inferior nasal turbinate and incubated with neopterin dissolved in either aqua destillata or in sodium chloride (0.9%) at the given amounts for 2, 5, 10, and 20 min. In a preliminary set of experiments, neopterin inhibited CBF following the 5 and 10 min. incubation protocol, respectively. However, analyses of pooled data from 28 experiments using sodium chloride (0.9%) as solvent could not confirm these findings: in no setting a significant reduction of CBF was detected at the end of the 20 min. experimental procedure. Therefore, we conclude that neopterin most likely does not affect the mucociliary clearance defense mechanisms in the course of infections of the nasal respiratory tract.

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Intracellular Cl− Regulation of Ciliary Beating in Ciliated Human Nasal Epithelial Cells: Frequency and Distance of Ciliary Beating Observed by High-Speed Video Microscopy

International Journal of Molecular Sciences ◽

10.3390/ijms21114052 ◽

2020 ◽

Vol 21 (11) ◽

pp. 4052

Author(s):

Makoto Yasuda ◽

Taka-aki Inui ◽

Shigeru Hirano ◽

Shinji Asano ◽

Tomonori Okazaki ◽

...

Keyword(s):

Epithelial Cells ◽

High Speed ◽

Beat Frequency ◽

Ciliary Beat Frequency ◽

Video Microscopy ◽

Nasal Epithelial Cells ◽

Ciliary Beating ◽

High Speed Video ◽

Human Nasal Epithelial Cells ◽

Ciliary Beat

Small inhaled particles, which are entrapped by the mucous layer that is maintained by mucous secretion via mucin exocytosis and fluid secretion, are removed from the nasal cavity by beating cilia. The functional activities of beating cilia are assessed by their frequency and the amplitude. Nasal ciliary beating is controlled by intracellular ions (Ca2+, H+ and Cl−), and is enhanced by a decreased concentration of intracellular Cl− ([Cl−]i) in ciliated human nasal epithelial cells (cHNECs) in primary culture, which increases the ciliary beat amplitude. A novel method to measure both ciliary beat frequency (CBF) and ciliary beat distance (CBD, an index of ciliary beat amplitude) in cHNECs has been developed using high-speed video microscopy, which revealed that a decrease in [Cl−]i increased CBD, but not CBF, and an increase in [Cl−]i decreased both CBD and CBF. Thus, [Cl−]i inhibits ciliary beating in cHNECs, suggesting that axonemal structures controlling CBD and CBF may have Cl− sensors and be regulated by [Cl−]i. These observations indicate that the activation of Cl− secretion stimulates ciliary beating (increased CBD) mediated via a decrease in [Cl−]i in cHNECs. Thus, [Cl−]i is critical for controlling ciliary beating in cHNECs. This review introduces the concept of Cl− regulation of ciliary beating in cHNECs.

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Daidzein-Stimulated Increase in the Ciliary Beating Amplitude via an [Cl−]i Decrease in Ciliated Human Nasal Epithelial Cells

International Journal of Molecular Sciences ◽

10.3390/ijms19123754 ◽

2018 ◽

Vol 19 (12) ◽

pp. 3754 ◽

Cited By ~ 4

Author(s):

Taka-aki Inui ◽

Makoto Yasuda ◽

Shigeru Hirano ◽

Yukiko Ikeuchi ◽

Haruka Kogiso ◽

...

Keyword(s):

Epithelial Cells ◽

Beat Frequency ◽

Cell Sheet ◽

Ciliary Beat Frequency ◽

Dependent Manner ◽

Nasal Epithelial Cells ◽

Ciliary Beating ◽

Human Nasal Epithelial Cells ◽

Cl Channels ◽

Ciliary Beat

The effects of the isoflavone daidzein on the ciliary beat distance (CBD, which is a parameter assessing the amplitude of ciliary beating) and the ciliary beat frequency (CBF) were examined in ciliated human nasal epithelial cells (cHNECs) in primary culture. Daidzein decreased [Cl−]i and enhanced CBD in cHNECs. The CBD increase that was stimulated by daidzein was mimicked by Cl−-free NO3− solution and bumetanide (an inhibitor of Na+/K+/2Cl− cotransport), both of which decreased [Cl−]i. Moreover, the CBD increase was inhibited by 5-Nitro-2-(3-phenylpropylamino)benzoic acid (NPPB, a Cl− channel blocker), which increased [Cl−]i. CBF was also decreased by NPPB. The rate of [Cl−]i decrease evoked by Cl−-free NO3− solution was enhanced by daidzein. These results suggest that daidzein activates Cl− channels in cHNECs. Moreover, daidzein enhanced the microbead transport driven by beating cilia in the cell sheet of cHNECs, suggesting that an increase in CBD enhances ciliary transport. An [Cl−]i decrease enhanced CBD, but not CBF, in cHNECs at 37 °C, although it enhanced both at 25 °C. Intracellular Cl− affects both CBD and CBF in a temperature-dependent manner. In conclusion, daidzein, which activates Cl− channels to decrease [Cl−]i, stimulated CBD increase in cHNECs at 37 °C. CBD is a crucial factor that can increase ciliary transport in the airways under physiological conditions.

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