Immune Netw.  2014 Apr;14(2):107-115. 10.4110/in.2014.14.2.107.

Phellinus linteus Extract Exerts Anti-asthmatic Effects by Suppressing NF-kappaB and p38 MAPK Activity in an OVA-induced Mouse Model of Asthma

• Yan GH
• Choi YH

• KMID: 2168021
• DOI: http://doi.org/10.4110/in.2014.14.2.107

Abstract

Phellinus linteus has been used as a traditional herbal medicine in Asian countries and is known to have anti-tumor, immunomodulatory, anti-inflammatory, and anti-allergic activities. However, the protective effects of P. linteus against experimental asthma have not been fully investigated. The objective of this study was to determine whether P. linteus ethanol extract (PLE) suppresses inflammatory response in an OVA-induced asthma model. As expected, the oral administration of PLE significantly inhibited eosinophilic airway inflammation and airway hyperresponsiveness in OVA-challenged BALB/c mice. Supporting these data, the augmentation of Th2 cytokines (IL-4, IL-5, and IL-13), eotaxin, and adhesion molecules in lung tissues and bronchoalveolar lavage fluid after OVA inhalation was markedly attenuated by PLE. Furthermore, PLE reduced OVA-induced activation of NF-kappaB and p38 MAPK in lung tissues. Therefore, our results suggest the potential of P. linteus as a therapeutic agent for asthma.

Keyword

Phellinus linteus; Asthma; Allergic inflammation; Th2 cytokine; NF-kappaB; p38

MeSH Terms

Administration, Oral
Animals
Asian Continental Ancestry Group
Asthma*
Bronchoalveolar Lavage Fluid
Cytokines
Eosinophils
Ethanol
Herbal Medicine
Humans
Inflammation
Inhalation
Interleukin-5
Lung
Mice*
NF-kappa B*
Ovum
p38 Mitogen-Activated Protein Kinases*
Cytokines
Ethanol
Interleukin-5
NF-kappa B
p38 Mitogen-Activated Protein Kinases

Figure

• 

  Figure 1 Airway hyperresponsiveness, differential cell counts in bronchoalveolar lavage (BAL) fluids, and histological evaluation of lung inflammation after OVA sensitization and treatment with Phellinus linteus ethanol extract (PLE). (A) All animals were nebulized with various concentrations of methacholine as a bronchoconstrictor. Data are shown as the percentage increase in Penh over the baseline, where the baseline Penh of the saline-treated control group is expressed as 100%. (B) Analysis of the effect of PLE, SB 239063, or BAY 11-7085 on OVA-induced differential cell counts in BAL fluid. EOS, eosinophil; NEU, neutrophil; MAC, macrophage; LYM, lymphocyte. (C) Paraffin-embedded lung sections were stained with H&E and periodic acid-Schiff (PAS). Magnification 200×. Bars indicate 50 µm. Data represent three independent experiments. (D) Inflammation scores. Total lung inflammation was defined as the average of the peribronchial and perivascular inflammation scores. (E) Quantitation of airway mucus expression. Sampling was performed 48 h after the last OVA challenge in mice. Results from three independent experiments with seven mice/group are given as means±SEM. SAL+SAL, saline-challenged mice administered saline; OVA+SAL, OVA-challenged mice administered saline; OVA+PLE, OVA-challenged mice administered PLE; OVA+SB 239063, OVA-challenged mice administered SB 239063; OVA+BAY, OVA-challenged mice administered BAY 11-7085. #p<0.05 vs. SAL+SAL; *p<0.05 vs. OVA+SAL.

• 

  Figure 2 Assessment of IgE, pro-inflammatory and Th2 cytokines, and adhesion molecules in the BAL fluids of OVA-sensitized mice treated with PLE. The levels of IgE, pro-inflammatory (TNF-α and IL-1β) and Th2 (IL-4, IL-5, and IL-13) cytokines, and adhesion molecules (eotaxin, ICAM-1, and VCAM-1) were quantified with ELISA. Sampling was performed 48 h after the last OVA challenge. Results from three independent experiments with seven mice/group are given as means±SEM. SAL+SAL, saline-challenged mice administered saline; OVA +SAL, OVA-challenged mice administered saline, OVA+PLE, OVA-challenged mice administered PLE; OVA+BAY, OVA-challenged mice administered BAY 11-7085. #p<0.05 vs. SAL+SAL; *p<0.05 vs. OVA+SAL.

• 

  Figure 3 Effect of PLE on OVA-induced NF-κB activation. The translocation of p65 to the nucleus (A) as well as IκB-α phosphorylation and degradation in cytoplasm (B) and NF-κB DNA binding activity (C) were assessed with western blot and EMSA, respectively. Density ratio vs. β-actin was measured using a densitometer. Results from three independent experiments with seven mice/group are given as means±SEM. SAL+SAL, saline-challenged mice administered saline; OVA +SAL, OVA-challenged mice administered saline; OVA+PLE, OVA-challenged mice administered PLE; OVA+SB 239063, OVA-challenged mice administered SB 239063. #p<0.05 vs. SAL+SAL; *p<0.05 vs. OVA+SAL.

• 

  Figure 4 Effect of PLE on OVA-induced p38 mitogen-activate protein kinase (MAPK) activation. Protein expressions of phosphorylated (p)-p38 and p38 in lung tissues were evaluated 48 h after the last OVA challenge. Density ratio vs. β-actin was measured using a densitometer. Results from three independent experiments with seven mice/group are given as means±SEM. SAL+SAL, saline-challenged mice administered saline; OVA+SAL, OVA-challenged mice administered saline; OVA+PLE, OVA-challenged mice administered PLE; OVA+SB 239063, OVA-challenged mice administered SB 239063. #p<0.05 vs. SAL+SAL; *p<0.05 vs. OVA+SAL.

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