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Key Event Title
Cilia Beat Frequency, Decreased
|Level of Biological Organization|
|multi-ciliated epithelial cell|
Key Event Components
|Abnormal ciliary motility||motile cilium||occurrence|
Key Event Overview
AOPs Including This Key Event
|AOP Name||Role of event in AOP||Point of Contact||Author Status||OECD Status|
|Oxidative stress [MIE] Leading to Decreased Lung Function [AO]||KeyEvent||Karsta Luettich (send email)||Open for comment. Do not cite|
|Ox stress-mediated CFTR/ASL/CBF/MCC impairment||KeyEvent||Karsta Luettich (send email)||Under development: Not open for comment. Do not cite|
|Homo sapiens||Homo sapiens||High||NCBI|
|Mus musculus||Mus musculus||High||NCBI|
|Rattus norvegicus||Rattus norvegicus||Moderate||NCBI|
|Oryctolagus cuniculus||Oryctolagus cuniculus||High||NCBI|
|Bos taurus||Bos taurus||High||NCBI|
|Cavia porcellus||Cavia porcellus||Moderate||NCBI|
|Lithobates catesbeianus||Rana catesbeiana||High||NCBI|
|All life stages||High|
Key Event Description
Cohesive beating of cilia lining the upper and lower respiratory tract is critical for efficient MCC. CBF is influenced by several factors including changes in the physical and chemical properties of the ASL (especially the periciliary fluid), structural modulation in the cilia, concentration of cyclic nucleotides cAMP and cGMP, and intracellular calcium (Ca2+). Formation of cyclic nucleotides such as cGMP is mediated by nitric oxide (NO), which is released by an enzyme family of nitric oxide synthases (NOSs) when the substrate L-arginine (L-Arg) is transformed to L-citrulline. NO activates its receptor protein, soluble guanylate cyclase (sGC), which catalyzes formation of cGMP from guanosine triphosphate (GTP). cGMP then activates protein kinase G (PKG) which has been implicated in the regulation of CBF (Jiao et al., 2011; Li et al., 2000). NO-dependent stimulation of CBF has also been associated with an increase in cAMP-dependent protein kinase A (PKA) (Di Benedetto et al., 1991; Lansley et al., 1992; Salathe et al., 1993; Sanderson and Dirksen, 1989; Schmid et al., 2007; Sisson et al., 1999; Uzlaner and Priel, 1999). An increase in intracellular endogenous cAMP was observed after treatment with isobutyl-1-methylxanthine that also increased CBF (Tamaoki et al., 1989). cAMP accumulation in the airway cilia has been shown to be dependent on Ca2+–calmodulin-dependent PDE1A and indirectly regulates CBF (Kogiso et al., 2018). Increase in CBF after treatment with NO substrate, L-arginine and inhibition of CBF by a NOS inhibitor, N-omega-nitro-L-arginine methyl ester (L-NAME) further provides evidence for the role of NO in increasing CBF (Jiao J. et al., 2011; Sisson J. H., 1995; Uzlaner and Priel, 1999; Yang et al., 1997). Modulation of CBF is not always accompanied by changes in cAMP levels. PKC activators, phorbol 12-myristate 13-acetate and L-o~-dioctanoylglycerol have been shown to decrease CBF in a concentration- and time-dependent manner in rabbit tracheal epithelial cells (Kobayashi et al., 1989). CBF has been shown to decrease after exposure to inhaled oxidants such as cigarette smoke across different species. A study with 120 subjects showed a significant decrease in nasal CBF following exposure to tobacco smoke (Agius et al., 1998). Exposure to cigarette smoke extract lead to reduction in forskolin-induced CBF in human sinonasal epithelium (Cohen et al., 2009) and isoproterenol- and methacholine-induced CBF in human adenoid tissues (Wang et al., 2012). This decrease in CBF and unresponsiveness to beta-agonist stimulation occurs in parallel to PKC activation and has been shown to be dependent on the duration of exposure to cigarette smoke in mice (Simet et al., 2010). Normal human bronchial epithelial cells exposed to aerosolized nicotine showed decreased CFTR and BK conductance, impaired CBF, ASL volume, and decreased expression of FOXJ1 and KCNMA1 (Garcia-Arcos et al., 2016). A concentration-dependent decrease in CBF has been observed after treatment with aldehydes. For example inhibition of cilia ATPase activity was observed after treatment with acetaldehyde, in ciliated bovine bronchial epithelial cells (Sisson et al., 1991). Acrolein, an aldehyde in the gas phase of cigarette smoke, induced ciliostasis at high concentrations (> 1 mM), after 5 min of treatment, and cellular necrosis after 3 hr. However, at lower concentrations (from 0.5‒1 mM), acrolein transiently reduced the CBF to 4 Hz (Romet et al., 1990).
How It Is Measured or Detected
There is no standardized method for measuring CBF. Digital high-speed video imaging with a manual count of CBF in slow motion video play is the most commonly used method for CBF measurement (Kim et al., 2011; Peabody et al., 2018). Photometry and video-microscopy have been used to measure CBF in vitro and ex vivo, including in ciliated bovine bronchial epithelial cells (Allen-Gipson et al., 2011; Sisson et al., 2003; Uzlaner and Priel, 1999), normal human bronchial epithelial cells (Feriani et al., 2017), human nasal epithelial cells (Dimova et al., 2005; Min et al., 1999b), human nasal ciliated epithelium (nasal brushings) (Agius et al., 1998), and mouse tracheal rings (Simet et al., 2010). CBF measurement in vitro generally involves mounting the tissue at the air-liquid interface on a stage followed by microscopic analysis and acquisition of images and/or video recordings of beating cilia. For in vivo and ex vivo measurements, Doppler optical coherence tomography (D-OCT) can also be applied, a mesoscopic non-contact imaging modality that provides high-resolution tomographic images and detects micromotion simultaneously (Jing et al., 2017). D-OCT has been used to quantitatively measure CBF in ex vivo rabbit tracheal cultures (Lemieux et al., 2015).
Domain of Applicability
Age-dependent decreases in CBF have been demonstrated in several species (e.g. guinea pigs, mice, and human) (Bailey et al., 2014; Grubb et al., 2016; Ho et al., 2001; Joki and Saano, 1997; Paul et al., 2013). In a study with 46 healthy subjects with a wide age distribution (mean 42, range 19–81 years), age was found to be negatively associated with airway clearance of inhaled 6-μm Teflon particles (Svartengren et al., 2005).
Female hormones, i.e. progesterone and estrogen, have been shown to have direct effect on CBF, i.e., progesterone reduces CBF, 17β-estradiol and progesterone receptor antagonists counteract progesterone effects, but estradiol alone has also been shown to have no effect on CBF. However, the mechanism by which these hormones modulate CBF is yet to be elucidated (Jain et al., 2012; Jia et al., 2011).
Evidence for Perturbation by Stressor
Treatment of human sinonasal epithelial cells with cigarette smoke condensate for 3 minutes significantly reduced forskolin-stimulated CBF (Cohen et al., 2009). CBF was also decreased in differentiated normal human bronchial epithelial cells exposed to whole cigarette smoke (Schmid et al., 2015), in cilia-bearing explant adenoid tissues treated with 5 and 10% cigarette smoke extract (Wang et al., 2012), in hamster oviducts treated various mainstream cigarette smoke fractions (Knoll et al., 1995), and in nasal epithelial cells fom smokers with moderate and severe chronic obstructive pulmonary disease (COPD) (Yaghi et al., 2012).
A concentration-dependent decrease in CBF has been observed after treatment with aldehydes. For example inhibition of cilia ATPase activity was observed after treatment with acetaldehyde, in ciliated bovine bronchial epithelial cells (Sisson et al., 1991).
Acrolein, an aldehyde in the gas phase of cigarette smoke, induced ciliostasis at high concentrations (> 1 mM), after 5 min of treatment, and cellular necrosis after 3 hr. However, at lower concentrations (from 0.5‒1 mM), acrolein transiently reduced the CBF to 4 Hz (Romet et al., 1990).
Normal human bronchial epithelial cells exposed to aerosolized nicotine showed decreased CFTR and BK conductance, CBF, ASL volume, and decreased expression of FOXJ1 and KCNMA1 (Garcia-Arcos et al., 2016).
Continuous, exposure of human nasal epithelial cells to different concentrations of ozone at 37°C for up to 4 weeks slightly (but not significantly) reduced CBF in healthy mucosa (7.1% at 500 µg/m3 and 10.3% at 1000 µg/m3), and significantly in chronically inflamed mucosa (20.5/16.4%) at 2 weeks. During the third and fourth week of exposure at these higher concentrations CBF was significantly reduced in both healthy (after 3 weeks: 18.7/37.5%; after 4 weeks: 11.1/33.3%) and chronically inflamed mucosa (after 3 weeks: 33.8/26.8%; after 4 weeks: 21.4/38.6%). Low ozone concentrations (100 µg/m3) appeared to not have an effect on CBF (Gosepath et al., 2000).
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