Allergy Asthma Immunol Res.  2014 Nov;6(6):548-557. 10.4168/aair.2014.6.6.548.

Oral Lovastatin Attenuates Airway Inflammation and Mucus Secretion in Ovalbumin-Induced Murine Model of Asthma

• Liou CJ
• Cheng PY
• Huang WC
• Chan CC
• Chen MC
• Kuo ML
• Shen JJ

• KMID: 2260178
• DOI: http://doi.org/10.4168/aair.2014.6.6.548

Abstract

PURPOSE
Lovastatin is an effective inhibitor of cholesterol synthesis. A previous study demonstrated that lovastatin can also suppress airway hyperresponsiveness (AHR) in murine model of asthma. We aimed to investigate the effect of lovastatin on mucus secretion and inflammation-associated gene expression in the lungs of murine model of asthma.
METHODS
Female BALB/c mice were sensitized and challenged with ovalbumin (OVA) by intraperitoneal injection, and orally administered lovastatin from days 14 to 27 post-injection. Gene expression in lung tissues was analyzed using real-time polymerase chain reaction. AHR and goblet cell hyperplasia were also examined. BEAS-2B human bronchial epithelial cells were used to evaluate the effect of lovastatin on the expression of cell adhesion molecules, chemokines, and proinflammatory cytokines in vitro.
RESULTS
We showed that lovastatin inhibits the expression of Th2-associated genes, including eotaxins and adhesion molecules, in the lungs of murine model of asthma. Mucin 5AC expression, eosinophil infiltration and goblet cell hyperplasia were significantly decreased in the lung tissue of murine model of asthma treated with lovastatin. Furthermore, lovastatin inhibited AHR and expression of Th2-associated cytokines in bronchoalveolar lavage fluid. However, a high dose (40 mg/kg) of lovastatin was required to decrease specific IgE to OVA levels in serum, and suppress the expression of Th2-associated cytokines in splenocytes. Activated BEAS-2B cells treated with lovastatin exhibited reduced IL-6, eotaxins (CCL11 and CCL24), and intercellular adhesion molecule-1 protein expression. Consistent with this, lovastatin also suppressed the ability of HL-60 cells to adhere to inflammatory BEAS-2B cells.
CONCLUSIONS
These data suggest that lovastatin suppresses mucus secretion and airway inflammation by inhibiting the production of eotaxins and Th2 cytokines in murine model of asthma.

Keyword

Asthma; cytokine; eosinophil; eotaxin; lovastatin; MUC5 AC

MeSH Terms

Animals
Asthma*
Bronchoalveolar Lavage Fluid
Cell Adhesion Molecules
Chemokines
Cholesterol
Cytokines
Eosinophils
Epithelial Cells
Female
Gene Expression
Goblet Cells
HL-60 Cells
Humans
Hyperplasia
Immunoglobulin E
Inflammation*
Injections, Intraperitoneal
Intercellular Adhesion Molecule-1
Interleukin-6
Lovastatin*
Lung
Mice
Mucin 5AC
Mucus*
Ovalbumin
Ovum
Real-Time Polymerase Chain Reaction
Cell Adhesion Molecules
Chemokines
Cholesterol
Cytokines
Immunoglobulin E
Intercellular Adhesion Molecule-1
Interleukin-6
Lovastatin
Mucin 5AC
Ovalbumin

Figure

• 

  Fig. 1 Effects of lovastatin on goblet cell hyperplasia in lung tissue. Lung sections were stained with periodic acid-Schiff (PAS) stain to assessgoblet cell hyperplasia in normal (A), OVA (B), L10 (C), and L40 (D) groups (200× magnification). The number of PAS-positive cells per 100 µm of basement membrane is visualized graphically (E). Data are presented as means ± SEM. **P<0.01 compared to the OVA group. Arrows indicate areas with gobletcell metaplasia.

• 

  Fig. 2 Effects of lovastatin on MUC5AC expression in the lungs. MUC5AC expression was analyzed by immunohistochemical staining(brown regions indicate positive staining; arrows) in normal (A), OVA (B), L10 (C), and L40 (D) groups (200× magnification). Results are expressed as the number of MUC5AC-positive cells per 100 µm (E). Data are presented as means ± SEM. *P<0.05 compared to the OVA group. **P<0.01 compared to the OVA group.

• 

  Fig. 3 Effects of lovastatin on the expression of MUC5AC, cytokines, chemokines, and adhesion molecules in the lung. CCL11 (A), CCL24 (B), ICAM-1 (C), MUC5AC (D), IFN-γ (E), IL-6 (F), IL-4 (G), IL-5 (H), and IL-13 (I) gene expression levels were determined using real-time RT-PCR. Data are presented as means ± SEM. *P<0.05 compared with OVA mice. **P<0.01 compared with OVA mice.

• 

  Fig. 4 Effects of lovastatin on eosinophil infiltration in lung tissue. Hematoxylin and eosin staining of lung tissue from normal (A), OVA (B), L10 (C), and L40 (D) groups (200×magnification). Arrows indicate areas with peribronchial and perivascular eosinophils.

• 

  Fig. 5 Effects of lovastatin on cytokine and chemokine levels in BEAS-2B cells. IL-6 (A), ICAM-1 (B), CCL11 (C), and CCL24 (D) levels were measured using ELISA. Data are presented as means ± SEM. *P<0.05 compared with the activated control group. **P<0.01 compared with the activated control group.

• 

  Fig. 6 Lovastatin suppression of eosinophil adhesion to BEAS-2B cells. HL-60 cells were treated with calcein AM and applied to BEAS-2B cell cultures. Results were observed using fluorescence microscopy. Note the adherence of HL-60 cells to normal (A) and TNF-α-activated BEAS-2B cells (B). Pretreatment of HL-60 cells with 10 µM (C), 20 µM (D), and 40 µM (E) of lovastatin reduced the adherence of HL-60 cells to BEAS-2B cells in a dose-dependent manner.

• 

  Fig. 7 Effects of lovastatin on bronchoalveolar lavage cell counts and airway hyperresponsiveness. (A) Both total cell number and specific cell counts were calculated for BALF samples. (B) The percentage of eosinophil cells in BALF. (C) AHR (shown as Penh values) was measured after mice were administered an inhalation dose of 10-40 mg/mL methacholine. Data are presented as means ± SEM. *P<0.05 compared with OVA mice. **P<0.01 compared with OVA mice.

• 

  Fig. 8 Lovastatin modulation of OVA-specific antibodies in serum. Levels of OVA-IgE (A), OVA-IgG1 (B), OVA-IgG2a (C), and cholesterol (D) in serum are shown. Lovastatin also modulates cytokine production by OVA-activated splenocytes: IL-4 (E), IL-5 (F), and IL-13 (G). Data are presented as means ± SEM. *P<0.05 compared with OVA mice. **P<0.01 compared with OVA mice.

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