Korean J Physiol Pharmacol.  2018 Jan;22(1):81-89. 10.4196/kjpp.2018.22.1.81.

Inhibitory effects of 2,6-di-tert-butyl-4-hydroxymethylphenol on asthmatic responses to ovalbumin challenge in conscious guinea pigs

• Jeong SY
• Lee JY

• KMID: 2398558
• DOI: http://doi.org/10.4196/kjpp.2018.22.1.81

Abstract

This study evaluated the anti-asthmatic activities of 2,6-di-tert-butyl-4-hydroxymethylphenol (DBHP) that is a potent phenolic antioxidant in edible vegetable oil. The effects of DBHP on bronchial asthma were evaluated by determining the specific airway resistance (sRaw) and tidal volume (TV) during the immediate asthmatic response (IAR) and the late-phase asthmatic response (LAR) in guinea pigs with aerosolized ovalbumin-induced asthma. Recruitment of leukocytes and the levels of biochemical inflammatory mediators were determined in the bronchoalveolar lavage fluids (BALFs), and histopathological surveys performed in lung tissues. DBHP significantly inhibited the increased sRaw and improved the decreased TV on IAR and LAR, and also inhibited recruitment of eosinophils and neutrophils into the lung, and release of biochemical inflammatory mediators such as histamine and phospholipase Aâ‚‚ from these infiltrated leukocytes, and improved pathological changes. However, anti-asthmatic activities of DBHP at oral doses of 12.5 to 50 mg/kg was less than those of dexamethasone (5 mg/kg, p.o.) and cromoglycate (10 mg/kg, p.o.), but more potent or similar to that of salbutamol (5 mg/kg, p.o.). These results in the present study suggest that anti-asthmatic effects of DBHP in the guinea pigs model of OVA-induced asthmatic responses principally are mediated by inhibiting the recruitments of the leukocytes and the release of biochemical inflammatory mediators from these infiltrated leukocytes.

Keyword

2,6-di-tert-butyl-4-hydroxymethylphenol; Immediate asthmatic response; Late-phase asthmatic response; PLAâ‚‚; sRaw

MeSH Terms

Airway Resistance
Albuterol
Animals
Asthma
Bronchoalveolar Lavage Fluid
Cromolyn Sodium
Dexamethasone
Eosinophils
Guinea Pigs*
Guinea*
Histamine
Leukocytes
Lung
Neutrophils
Ovalbumin*
Phenol
Phospholipases
Tidal Volume
Vegetables
Albuterol
Cromolyn Sodium
Dexamethasone
Histamine
Ovalbumin
Phenol
Phospholipases

Figure

• 

  Fig. 1 Chemical structure of 2,6-di-tert-bytyl-4-hydroxymethylphenol (A) and diagram of the double-chambered plethysmograph (B) used in this experiment.This double-chambered plethysmograph is a basic method for measurement of the specific airway resistance (sRaw) together with the standard parameters tidal volume and respiratory rate in conscious animals placed into the plethysmograph box (HSE type 855, Hugo Sachs Elektronik, Germany). The phase shift between two the nasal and thoracic respiratory flows is measured with two differential pressure transducers (PT5, Grass Instument Co., USA). The PULMODYN “PENNOCK 89” software is able to record the signals and calculate the sRaw with a respiratory analyzer (7E polygraph, Grass Instument Co., USA), and the PLUGSYS 603 system is used to control the valves from the plethysmograph boxes and interface the boxes to the computer (This diagram modified from an original kindly provided by Hugo Sachs Elektronik company, Germany).

• 

  Fig. 2 Effect of DBHP on specific airway resistance (sRaw) to the antigen challenge in IAR (A) and LAR (B).Test drugs were orally administered 1 h prior to OVA challenge, additionally 12 h later. Percentage increase of sRaw (%)=[(sRaw after OVA challenge/sRaw before OVA challenge)−1]×100. Each value represents the mean±SEM (n=6). Values are statistically significant at #p<0.05 compared with the vehicle control; *p<0.05 and **p<0.01 compared with OVA control. DBHP, 2,6-di-tert-butyl-4-hydroxymethylphenol; DEX, dexamethasone; CRO, cromoglycate; SAL, salbutamol.

• 

  Fig. 3 Effect of DBHP on the tidal volume (TV) to the antigen challenge in IAR (A) and LAR (B).Test drugs were orally administered 1 h prior to OVA challenge, additionally 12 h later. Percentage decrease of TV (%)=[1−(TV after OVA challenge/TV before OVA challenge)]×100. Each value represents the mean±SEM (n=6). Values are statistically significant at #p<0.05 compared with the vehicle control; *p<0.05 and **p<0.01 compared with OVA control. DBHP, 2,6-di-tert-butyl-4-hydroxymethylphenol; DEX, dexamethasone; CRO, cromoglycate; SAL, salbutamol.

• 

  Fig. 4 Effect of DBHP on the recruitment of leukocytes in BALF.The number of total leukocytes (A), eosinophils (B), and neutrophils (C) were counted in BALF. Test drugs were orally administered 1 h prior to OVA challenge, additionally 12 h later. Each value represents the mean±SEM (n=6). Values are statistically significant at #p<0.05 compared with the vehicle control; *p<0.05 and **p<0.01 compared with OVA control. DBHP, 2,6-di-tert-butyl-4-hydroxymethylphenol; DEX, dexamethasone; CRO, cromoglycate; SAL, salbutamol.

• 

  Fig. 5 Effect of DBHP on the release of biochemical mediators in BALF.Histamine content (A), protein exudates (B), and the specific activity of PLA2 (C) were measured fluorometrically and colorimetrically in BALF. Test drugs were orally administered 1 h prior to OVA challenge, additionally 12 h later. Each value represents the mean±SEM (n=6). Values are statistically significant at #p<0.05 compared with the vehicle control; *p<0.05 and **p<0.01 compared with OVA control. DBHP, 2,6-di-tert-butyl-4-hydroxymethylphenol; DEX, dexamethasone; CRO, cromoglycate; SAL, salbutamol.

• 

  Fig. 6 Effect of DBHP on the histopathological survey in lung tissue.Representative images of the H&E-stained sections of lung tissue from; (A) vehicle control, showed a normal architecture and non-infiltration of inflammatory cells. (B) OVA control, exhibited the recruitment of leukocytes in alveolar sac (black arrow), peripheral vasculature (red arrow) and terminal bronchioles (yellow arrow). (C) asthmatic lung tissue treated with DBHP (50 mg/kg, p.o.) at 1 h before, and 12 h after OVA challenge, was reduced the recruitment of leukocytes into alveolar sacs (pink arrow). (D) asthmatic lung tissue treated with dexamethasone (5 mg/kg, p.o.) at 1 h before, and 12 h after OVA challenge, was also improved pathological changes with mild infiltration of leukocytes around bronchioles (purple arrow) (original magnification; ×100).

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