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. 2021 Jan 8;6(1):34-41.
doi: 10.1002/lio2.518. eCollection 2021 Feb.

Effects of narrow-band UVB on nasal symptom and upregulation of histamine H1 receptor mRNA in allergic rhinitis model rats

Seiichiro Kamimura  1 , Yoshiaki Kitamura  1 , Tatsuya Fujii  1 , Kentaro Okamoto  2 , Nanae Sanada  2 , Natsuki Okajima  2 , Tomoharu Wakugawa  2 , Hiroyuki Fukui  1 , Hiroyuki Mizuguchi  3 , Noriaki Takeda  1
Affiliations

Effects of narrow-band UVB on nasal symptom and upregulation of histamine H1 receptor mRNA in allergic rhinitis model rats

Seiichiro Kamimura et al. Laryngoscope Investig Otolaryngol. .

Abstract

Background: Phototherapy with narrow-band ultraviolet B (narrow-band UVB) is clinically effective treatment for atopic dermatitis. In the present study, we examined the effects of intranasal irradiation with narrow-band UVB on nasal symptom, upregulation of histamine H1 receptor (H1R) gene expression and induction of DNA damage in the nasal mucosa of allergic rhinitis (AR) model rat.

Methods: AR model rats were intranasally irradiated with 310 nm of narrow-band UVB. Nasal mucosal levels of H1R mRNA were measured using real-time quantitative reverse transcriptase (RT)-PCR. DNA damage was evaluated using cyclobutane pyrimidine dimer (CPD) immunostaining.

Results: In toluene 2,4-diisocyanate (TDI)-sensitized rats, TDI provoked sneezes and H1R gene expression in the nasal mucosa. Intranasal pre-irradiation with 310 nm narrow-band UVB at doses of 600 and 1400, but not 200 mJ/cm2 significantly inhibited the number of sneezes and upregulation of H1R gene expression provoked by TDI. CPD-positive cells appeared in the nasal mucosa after intranasal narrow-band UVB irradiation at a dose of 1400, but not 200 and 600 mJ/cm2. The suppression of TDI-provoked sneezes and upregulation of H1R gene expression lasted 24 hours, but not 48 hours, after narrow-band UVB irradiation with a dose of 600 mJ/cm2.

Conclusions: Intranasal pre-irradiation with narrow-band UVB dose-dependently inhibited sneezes and upregulation of H1R gene expression of the nasal mucosa in AR model rats, suggesting that the inhibition of nasal upregulation of H1R gene expression suppressed nasal symptom. The suppression after narrow-band UVB irradiation at a dose of 600 mJ/cm2 was reversible without induction of DNA damage. These findings indicated that low-dose narrow-band UVB phototherapy could be effectively and safely used for AR treatment in a clinical setting.

Level of evidence: NA.

Keywords: allergic rhinitis; apoptosis; histamine H1 receptor; narrow‐band ultraviolet B; phototherapy.

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Conflict of interest statement

The authors have no conflicts of interest directly relevant to the content of this article.

Figures

FIGURE 1
FIGURE 1
Experimental protocol. Rats were sensitized with 10 μL of a 10% solution of TDI in ethyl acetate, which was applied to the bilateral nasal vestibule on day 1‐5 and day 8‐12. Narrow‐band UVB of 310 nm was irradiated intranasally at doses of 200, 600 and 1400 mJ/cm2 on day 22. Three hours after narrow‐band UVB irradiation, we applied TDI to the bilateral nasal vestibule again to provoke nasal symptom. Immediately after TDI provocation, the number of sneezes was counted during 10 minutes
FIGURE 2
FIGURE 2
Effects of intranasal irradiation with 310 nm narrow‐band UVB on TDI‐induced up‐regulation of H1R mRNA of the nasal mucosa, A, and sneezes, B, of TDI‐sensitized rats. Narrow‐band UVB at doses of 200, 600, and 1400 mJ/cm2 was irradiated intranasally through bilateral nostrils of rats. Three hours after narrow‐band UVB irradiation, TDI was applied again to provoke nasal symptom. Immediately after TDI provocation, the number of sneezes was counted during 10 minutes. The nasal mucosa of rats was dissected 4 hours after TDI provocation. We determined H1R mRNA level using real‐time PCR. Control rats were treated with vehicle. The data are expressed as means ± SEM. n = 9 (200 mJ/cm2), 8 (600 mJ/cm2), 7 (1400 mJ/cm2), 20 (TDI without irradiation), 20 (control). ## P < .01 vs control; *P < .05, **P < .01 vs TDI
FIGURE 3
FIGURE 3
Effects of intranasal irradiation with 310 nm narrow‐band UVB on induction of apoptosis in nasal mucosa cells of TDI‐sensitized rats. Narrow‐band UVB at doses of 200, 600, and 1400 mJ/cm2 was irradiated intranasally through bilateral nostrils of rats. Three hours after narrow‐band UVB irradiation, TDI was applied to the nasal vestibule to induce nasal symptom. Four hours after provocation, the nasal mucosa of rats was dissected, fixed in 4% paraformaldehyde, embedded in OCT compound, and rapidly frozen. The tissue was cut into 10 μm‐thick sections. The tissue sections were stained using DeadEnd Fluorometric TUNEL System, counter‐stained with propidium iodide and observed using a fluorescent microscope. Control rats were treated with vehicle. As a positive control, the nasal mucosa of control rats was treated with DNase. Apoptotic cells were indicated as green fluorescence and nuclei were indicated as red fluorescence. Scale bar indicates 50 μm
FIGURE 4
FIGURE 4
Effects of intranasal irradiation with 310 nm narrow‐band UVB on UV‐induced DNA damage in nasal mucosa cells of TDI‐sensitized rats. Narrow‐band UVB at doses of 200, 600, and 1400 mJ/cm2 was irradiated intranasally through bilateral nostrils of rats. Three hours after narrow‐band UVB irradiation, TDI was applied to the nasal vestibule to induce nasal symptom. Four hours after provocation, the nasal mucosa of rats was dissected, fixed in 4% paraformaldehyde, embedded in OCT compound, and rapidly frozen. The tissue was cut into 10 μm‐thick sections. The tissue sections were immunostained using anti‐CPD antibody, counter stained with propidium iodide, and observed using a fluorescent microscope. Control rats were treated with vehicle. CPD‐positive cells were indicated as green fluorescence, and nuclei were indicated as red fluorescence. Scale bar indicates 50 μm
FIGURE 5
FIGURE 5
Time course of TDI‐induced H1R mRNA upregulation in the nasal mucosa, A, and sneezes, B, after intranasal irradiation with 310 nm narrow‐band UVB of TDI sensitized rats. Narrow‐band UVB at a dose of 600 mJ/cm2 was irradiated intranasally through bilateral nostrils of rats. TDI provocation was performed 3 hours, 24 hours, and 48 hours later. Immediately after TDI provocation, we counted the number of sneezes during 10 minutes. Four hours after TDI provocation, the nasal mucosa was dissected. H1R gene expression level was determined using real‐time PCR. Control rats were treated with vehicle. The data are presented as means ± SEM. n = 3 (3 hours), 4 (24 hours), 3 (48 hours), 4 (TDI without irradiation), 4 (control). # P < .05, ## P < .01 vs control; *P < .05, **P < .01 vs TDI
FIGURE 6
FIGURE 6
Time course of induction of apoptosis, A, and DNA damage, B, in the nasal mucosa after intranasal irradiation with 310 nm narrow‐band UVB of TDI‐sensitized rats. Narrow‐band UVB at a dose of 600 mJ/cm2 was irradiated intranasally through bilateral nostrils of rats. TDI provocation was performed 3 hours, 24 hours, and 48 hours after narrow‐band UVB irradiation. Four hours after provocation, the nasal mucosa of rats was dissected fixed in 4% paraformaldehyde, embedded in OCT compound, and rapidly frozen. The tissue was cut into 10 μm‐thick sections. The tissue sections were stained using DeadEnd Fluorometric TUNEL System and anti‐CPD antibody, counter‐stained with propidium iodide, and observed using a fluorescent microscope. A, Apoptotic cells and B, CPD‐positive cells were indicated as green fluorescence, and nuclei were indicated as red fluorescence. Scale bar indicates 50 μm
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