J Rhinol.  2020 Nov;27(2):108-116. 10.18787/jr.2020.00325.

Bacillus clausii, a Foreshore-Derived Probiotic, Attenuates Allergic Airway Inflammation Through Downregulation of Hypoxia Signaling

• Park H
• Jung AY
• Chang CS
• Kim YH

• KMID: 2508955
• DOI: http://doi.org/10.18787/jr.2020.00325

Abstract

Background and Objectives
The immunomodulatory effects and mechanism of probiotics in allergic airway disease are largely unknown. We studied whether Bacillus clausii (BC), a probiotic derived from mudflats, had anti-allergic effects and compared the results with those of Lactobacillus paracasei (LP). We also examined whether the anti-allergic mechanisms of probiotics are associated with hypoxia signaling. Materials and Method: Forty-two BALB/c mice were randomly assigned to six experimental groups: controls, ovalbumin (OVA)-induced mice for inducing asthma, and OVA-induced mice that were orally administered LP or BC, at 1×10⁹ or 5×10⁹ CFU/mL each. We performed differential cell count testing on bronchoalveolar lavage fluid (BALF), lung histopathology, serum totals and OVA-specific IgE and IgG1 assessments, Th2 cytokine titers (IL-4, IL-5) in BALF and pulmonary parenchyma, quantitative PCR for heme oxygenase (HO)-1 and Hif-1α, and immunohistochemistry.
Results
Compared to the OVA group mice, OVA-sensitized mice treated with LP or BC showed significantly reduced numbers of eosinophils and neutrophils in the BALF (p<0.05). Both probiotics also significantly reduced pulmonary inflammation and eosinophil infiltration. Mice in the LP or BC group had a substantially lower titer of IL-4 and IL-5 in BALF, and decreased IL-4 and IL-5 expression in the lung parenchyma. Real-time PCR and immunohistochemistry showed that both LP and BC could significantly suppress HO-1 and Hif-1α expression in asthmatic mice (p<0.05).
Conclusion
BC can attenuate murine allergic asthma by regulating HIF-1α signaling, and its anti-allergic effect is comparable to that of LP.

Keyword

Asthma; Probiotics; Hypoxia-inducible factor 1; Heme oxygenase-1

Figure

• 

  Fig. 1. Time schedule of the asthma mouse model. Male BALB/c mice were sensitized intraperitoneally to OVA on day 0 and 14 and mice were challenged daily from day 28 until day 30 by intranasal instillation. Mice were treated for 38 days with either LP or BC by oral gavage. Mice in the LP1 or BC1 group were administered 0.2 mL of LP or BC at a dose of 1×109 CFU/mL. Mice in the LP5 or BC5 group were fed 0.2 mL of LP or BC, at a dose of 5×109 CFU/mL, respectively. OVA: ovalbumin, LP: Lactobacillus paracasei, BC: Bacillus clausii.

• 

  Fig. 2. Effects of probiotics on hematological and histological changes. BALF was collected 24 h after the last challenge. The lavage fluids were centrifuged, and cell pellets were counted using a hemocytometer after cell staining with trypan blue. A: BALF total cells. B: eosinophils. C: neutrophils, D: macrophages. E: lymphocytes. F: Lung tissues stained with H&E (×200), showing infiltration of inflammatory cells. G: Lung tissues stained with PAS (×200), showing airway goblet cells. The data represent results from seven mice per group. All data are expressed as mean±SD. P-values compared to the OVA group. OVA: ovalbumin. LP: Lactobacillus paracasei, BC: Bacillus clausii.

• 

  Fig. 3. Effects of probiotics on Th2 inflammation. A: Total IgE levels in the serum. B: Titers of plasma OVA-specific soluble IgE. C: OVAspecific IgG1 in the six groups. D: IL-4. E: IL-5 concentration in BALF was measured by ELISA. F: Cytokine IL-4. G: IL-5 mRNA levels in BALF were determined by quantitative RT-PCR. The data represent results from seven mice per group. All data are expressed as mean±SD. P-values compared to the OVA group. OVA: ovalbumin, LP: Lactobacillus paracasei, BC: Bacillus clausii.

• 

  Fig. 4. Effects of probiotics on hypoxia signaling. A: qRT-PCR was performed to measure the mRNA expression of HO-1 in lung tissue. B: qRT-PCR was performed to measure the mRNA expression of HIF-1α in lung tissue. The data represent results from seven mice per group. C: Immunohistochemical staining of HO-1 in lung tissues (×200 magnification). D: Immunohistochemical staining of HIF-1α in lung tissues (×400 magnification). Red arrows indicate HIF-1α-positive cells. All data are expressed as mean±SD. P-values compared to the OVA group. OVA: ovalbumin, LP: Lactobacillus paracasei. BC: Bacillus clausii.

Reference

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