[논문]Suppressive Effect of Carnosol on Ovalbumin-Induced Allergic Asthma

Lee, Jung-Eun; Im, Dong-Soon

doi:10.4062/biomolther.2020.050

[국내논문] Suppressive Effect of Carnosol on Ovalbumin-Induced Allergic Asthma 원문보기

Biomolecules & therapeutics, v.29 no.1, 2021년, pp.58 - 63

Lee, Jung-Eun (Laboratory of Pharmacology, College of Pharmacy, and Department of Life and Nanopharmaceutical Sciences, Graduate School, Kyung Hee University) , Im, Dong-Soon (Laboratory of Pharmacology, College of Pharmacy, and Department of Life and Nanopharmaceutical Sciences, Graduate School, Kyung Hee University)

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Asthma is a chronic obstructive lung disease characterized by recurrent episodes of bronchoconstriction and wheezing. Conventional asthma treatment involves the suppression of airway inflammation or improving airway flow. Rosmarinus officialis, also known as rosemary, is a Mediterranean plant that is used for the treatment of inflammatory diseases. Carnosol, a diterpenoid found in rosemary extracts, has been known to exhibit anti-inflammatory, anti-tumor, and anti-oxidant effects. The effect of carnosol on allergic responses has not been tested yet. The effect of carnosol on a murine allergic asthma model were investigated. Carnosol inhibited the degranulation of RBL-2H3 mast cells. Carnosol treatment inhibited the increase in the number of eosinophils in the bronchoalveolar lavage fluids (BALF) of mice treated with ovalbumin. Carnosol treatment also inhibited inflammatory responses and mucin production in histologic studies. Carnosol treatment inhibited the increases of IL-4 and IL-13 cytokines expression in both BALF and the lungs. These results suggest that carnosol may have a potential for allergic asthma therapy.

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표/그림 (5)

그림 Fig. 1. Effect of carnosol on β-hexosaminidase release in RBL- 2H3 cells. RBL-2H3 cells were incubated with anti-DNP IgE for 18 h, and challenged with DNP human serum albumin (HSA). Cells were treated with carnosol at the indicated concentrations 30 min before antigen challenge. Basal degranulation was calculated from the samples without IgE and HSA. The degranulation from samples treated with IgE and HSA was used as a positive control. The results are presented as means ± SEs of values from three independent experiments. ***p<0.001 vs. the non-HSA-treated group. ^#p<0.05, ^##p<0.01, ^###p<0.001 vs. the HSA-treated group.
그림 Fig. 2. Suppressive effects of carnosol on the increase in immune cells in BALF by OVA. (A) Mice were sensitized with OVA twice by i.p. injection on D0 and D14, and later challenged on D28, D29, and D30 with nebulized OVA. Carnosol was administrated intraperitoneally at the dose of 5 mg/kg, 30 min before OVA challenge. Cells in BALF were stained using May-Grünwald stain and counted. (B) Total cell counts, eosinophils, and macrophages in BALF. (C) Lymphocytes counts in the BALF. The results are presented the mean ± SE cell count values (n=5). ***p<0.001 vs. the PBS-treated group, ^###p<0.001 vs. the OVA-treated group.
그림 Fig. 3. Carnosol treatment inhibited the expression of cytokines at the mRNA level in the BALF cells and lungs. Analysis of mRNA expression of Th2 cytokines (IL-4, IL-5, and IL-13) and Th17 cytokine (IL-17A) from the BALF cells (A) or the lung tissues (B). Relative mRNA levels of cytokines were quantified as ratios to GAPDH transcript levels. Values represent the mean ± SE (n=5). ***p<0.001 vs. the PBStreated group, ^#p<0.05, ^##p<0.01, ^###p<0.001 vs. the OVA-treated group.
그림 Fig. 4. Protective effect of carnosol against airway inflammation and mucin production. (A) Panels show H&E-stained sections of lung tissues from the PBS group, OVA group, and carnosol plus OVA group. Sections from mice in the OVA-induced and carnosoltreated OVA-induced groups showed an accumulation of eosinophils (small navy blue dots, green arrows). (B) Panels show PASstained sections of lung tissues. Mucin is stained in purple color (red arrows). (C) Lung inflammation was semi-quantitatively evaluated. Histological findings were scored as described in the materials and methods section. (D) Mucous production was evaluated by counting the number of PAS-positive cells per mm of bronchiole (n=5 per group). Bar=50 μm. Graphs represent the mean ± SE values (n=5). ***p<0.001 vs. the PBS-treated group, ^###p<0.001 vs. the OVAtreated group.
그림 Fig. 5. Effect of carnosol treatment on IL-4 protein levels in BALF. ELISA was used to measure the protein levels of IL-4 in BALF (n=5 per group). The results represent the mean ± SE of protein levels (n=5). **p<0.01 vs. the PBS-treated group, ^##p<0.01 vs. the OVAtreated group.

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대상 데이터

Carnosol and other materials were purchased from Sigma- Aldrich (St. Louis, MO, USA).
Female five-week-old BALB/c mice were purchased from Daehan Biolink (Seoul, Korea). They were housed in the laboratory animal facility at Pusan National University and fed water and food ad libitum.
Biolink (Seoul, Korea). They were housed in the laboratory animal facility at Pusan National University and fed water and food ad libitum. The Pusan National University Institutional Animal Care Committee reviewed and approved the protocol with respect to ethical issues and scientific care (Ap- proval Number, ED-PNU2019-0011)

이론/모형

For statistical significance, analysis of variance (ANO- VA) was used, followed by Turkey’s post hoc test using Graph- Pad Prism software (GraphPad Software, Inc., La Jolla, CA, USA). p values<0.

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