Clin Exp Otorhinolaryngol.  2019 May;12(2):196-205. 10.21053/ceo.2019.00094.

Antiallergic Effect of Hizikia fusiformis in an Ovalbumin-Induced Allergic Rhinitis Mouse Model

• Zhang YL
• Shin HJ
• Lee JH
• Lee J

• KMID: 2447429
• DOI: http://doi.org/10.21053/ceo.2019.00094

Abstract

OBJECTIVES
The extract of Hizikia fusiformis is known to exhibit anticancer, antiatopic and antioxidant activities. We aimed to investigate the extract of H. fusiformis on allergic rhinitis inflammation in a mouse model.
METHODS
The 4-week-old BALB/c mice were randomly assigned into four groups: group A, control group (n=9); group B, allergic rhinitis group (n=10); group C (n=10) received 300 mg/kg of H. fusiformis during nasal challenging period; group D (n=10) received 600 mg/kg of H. fusiformis during general sensitization period and 300 mg/kg of H. fusiformis during nasal challenging period. Allergic inflammation was made with ovalbumin (OVA) and alum then challenged intranasally with OVA. H. fusiformis was intraperitoneally administered 3 hours before the OVA administration. Allergic symptom score and the levels of immunoglobulin G1 (IgG1), IgG2a, OVA-specific IgE antibodies, levels of cytokines in the nasal mucosa and in spleen cell culture supernatant, such as tumor necrosis factor alpha (TNF-α), interleukin 4 (IL-4), IL-5, IL-13, and IL-10 were assessed. The percentage of regulatory T cell was analyzed by flow cytometry. Eosinophilic infiltration and goblet cell hyperplasia were also evaluated.
RESULTS
H. fusiformis administered groups C and D showed significant inhibitory effects on nasal symptoms, IL-13 mRNA expression and eosinophil infiltration/goblet cell hyperplasia in the nasal tissue; OVA-specific IgE production in serum (P<0.05). In group D, H. fusiformis treatment downregulated IL-4, IL-5, IL-13, TNF-α, and IL-10 cytokine expression in splenocyte culture as well as significantly decreased IgG2a, IgG1 levels in serum compared with group B (P<0.05). However, the expressions of IL-5, interferon-γ and forkhead box P3 mRNA did not change in groups C and D.
CONCLUSION
H. fusiformis could induce antiallergic inflammation by suppressing the T-helper type 2 cytokine production (IL-13) locally and systemically, OVA-specific IgE formation, goblet cell hyperplasia, and eosinophilic infiltration in a mouse model of allergic rhinitis. Thus, H. fusiformis could be considered as a potential therapeutic agent in treating allergic rhinitis.

Keyword

Allergic Rhinitis; Th2 Cells; Inflammation; Mice

MeSH Terms

Animals
Antibodies
Cell Culture Techniques
Cytokines
Eosinophils
Flow Cytometry
Goblet Cells
Hyperplasia
Immunoglobulin E
Immunoglobulin G
Immunoglobulins
Inflammation
Interleukin-10
Interleukin-13
Interleukin-4
Interleukin-5
Mice*
Nasal Mucosa
Ovalbumin
Ovum
Rhinitis, Allergic*
RNA, Messenger
Spleen
Th2 Cells
Tumor Necrosis Factor-alpha
Antibodies
Cytokines
Immunoglobulin E
Immunoglobulin G
Immunoglobulins
Interleukin-10
Interleukin-13
Interleukin-4
Interleukin-5
Ovalbumin
RNA, Messenger
Tumor Necrosis Factor-alpha

Figure

• 

  Fig. 1. Schematic representation of the experimental protocol. (General sensitization) BALB/c mice were sensitized with 25 μg of ovalbumin (OVA) and 2 mg of aluminum hydroxide gel on days 0, 7, and 14. (Local sensitization) all mice except for group A received intranasal challenge with OVA from day 21 to 27. In group C, mice received Hizikia fusiformis (H. fusiformis, 300 mg/kg) 3 hours before intranasal OVA challenge from day 21 to 25. In group D, mice received H. fusiformis (600 mg/kg) 3 hours before intraperitoneal OVA administration on days 0, 7, and 14 and H. fusiformis (300 mg/kg) 3 hours before intranasal OVA challenge. PBS, phosphate-buffered saline; IP, intraperitoneal injection.

• 

  Fig. 2. Effect of Hizikia fusiformis on levels of (A) allergic symptom score and (B-D) serum ovalbumin (OVA)-specific immunoglobulins in each group. Administration of H. fusiformis suppressed allergic symptoms including rubbing and sneezing, and serum immunoglobulin E (IgE) (B), IgG1 (C), and IgG2a (D) levels. A, control group; B, allergic rhinitis group; C, H. fusiformis treatment group during nasal challenging period; D, H. fusiformis treatment group during general sensitization and nasal challenging period; NS, not significant. *P<0.05.

• 

  Fig. 3. Effect of Hizikia fusiformis on the (A) production of Th1/Th2 cytokines and (B) induction of regulatory T cell subgroups in spleen cell culture. (A) In spleen cell culture supernatant, interleukin 4 (IL-4), IL-5, IL-13, IL-10 and tumor necrosis factor alpha (TNF-α) concentrations were decreased significantly in group D compared with the group B. (B) The frequency of CD4+CD25+FOXP3+ was decreased significantly in group D when the splenocyte cultured to OVA compared to the positive control group B. However, others did not show significant change after treatment. Experiments were performed three times. A, control group; B, allergic rhinitis group; C, H. fusiformis treatment group during nasal challenging period; D, H. fusiformis treatment group during general sensitization and nasal challenging period; NS, not significant; SSC-H, side scatter height; FSC-H, forward scatter height; FITC, fluorescein isothiocyanate; APC, allophycocyanine; PE, phycoerythrin. *P<0.05.

• 

  Fig. 4. Effect of Hizikia fusiformis on the nasal tissue cytokine expression. Local cytokine mRNA expression levels in the nasal mucosa were evaluated using real-time polymerase chain reaction after H. fusiformis administration. Transcriptional activity of interleukin 13 (IL-13) decreased significantly (B); however, those of IL-5 (A), interferon γ (IFN-γ; C), and FOXP3 (D) did not significantly change in the groups C and D compared with the positive control group B. A, control group; B, allergic rhinitis group; C, H. fusiformis treatment group during nasal challenging period; D, H. fusiformis treatment group during general sensitization and nasal challenging period; NS, not significant. *P<0.05

• 

  Fig. 5. Histopathological findings. (A, B) Eosinophil infiltration in the nasal mucosa. (C, D) Goblet cell hyperplasia in the nasal epithelium. (A) Subepithelial inflammatory cell infiltration including eosinophil was evident in group B and decreased in Hizikia fusiformis treatment groups C and D (Sirius red stain, ×400; yellow arrows, eosinophils). (B) Eosinophil count was significantly reduced in treatment groups C and D compared with the positive group B. (C) Increased number of goblet cell in epithelial layer was identified in group B (black arrows, goblet cells). (D) Systemic administration of H. fusiformis suppress goblet cell hyperplasia in groups C and D compared with the positive control group B (Periodic acid-Schiff stain, ×400). A, control group; B, allergic rhinitis group; C, H. fusiformis treatment group during nasal challenging period; D, H. fusiformis treatment group during general sensitization and nasal challenging period; HPF, high power field. *P<0.05.

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