The effect of carbonyl-sulphide containing medicinal water bath treatment on LPS-induced airway inflammation in mice

The effect of carbonyl-sulphide containing medicinal water bath treatment on LPS-induced airway inflammation in mice

Conclusion:The carbonyl-sulphide-containing Harkany medicinal water attenuates inflammatory cell content in the mouse model of endotoxin-evoked subacute airway inflammation. Harkány medicinal water does not influence bronchial hyperreactivity after LPS administration in vivo.

Background: Carbonyl-sulphide is converted to hydrogen-sulphide (H

S) in the body, it stimulates capsaicin-sensitive sensory nerves and induces the release of sensory neuropeptides, therefore it is likely to modulate the inflammatory processes and neuro-immune interactions. The aim of the present study was to investigate the effect of this medicinal water bath on a pneumonitis model of the mouse.

Methods: Male CD1 mice were bathed in 37 ^oC medicinal water or the same temperature distilled water for 20 min/day, during a 14-day experimental period (Fig.1). Airway inflammation was induced with intranasal administration of E. coli (serotype 083) endotoxin (LPS).

Airway hyper-reactivity to inhaled carbachol was determined in conscious mice with the Buxco whole body plethysmograph (Fig.2.,3)before and 24 h after LPS administration. Bronchoalveolar lavage (BAL) was performed to determine the inflammatory cell profile of the airways with flow-cytometry (Fig. 4,5,6). The right lung of each animal was fixed in formaldehyde, embedded in paraffin and stained with hematoxylin and eosin for histopathological investigation (Fig. 8a,b,c). The left lung was frozen in liquid nitrogen for measuring the inflammatory cytokine IL-1β with ELISA from the homogenates (Fig.7).

Fig.1. Constant temperature

water bath Fig.2. Buxco whole body

plethysmograph

A Markovics

, A Kemeny

, A Perkecz

, L Kereskai

, Zs Helyes

, E Pinter

1Department of Pharmacology and Pharmacotherapy, ²Janos Szentagothay Research Center, ³Department of Pathology, University of Pecs, Hungary

Fig.8a,b,c. Representative histological pictures from the lungs of a)PBS-treated, b)distilled water treated, c)medicinal water treated mice (100×)

0 11 22

0 2 4 6 8 10 12

PBS control

LPS+distilled water

LPS+medicinal water

Concentration of carbachol (mM)

Mean Penh value

Fig.3. Medicinal water bath did not influence bronchial hyperreactivity to inhaled carbachol

(n=24/group, p 0.05)

distilled water medicinal water 0

2000 4000 6000 8000 10000 12000 14000 16000

Concentration of IL-1

pg/g wet tissue

distilled water medicinal water 0

500 1000 1500 2000 2500 3000 3500 4000

*

Lymphocytes

Cell content/ml fluid

distilled water medicinal water 0

200 400 600 800 1000 1200 1400

*

Granulocytes

Cell content/ml fluid

distilled water medicinal water 0

300 600 900 1200 1500 1800 2100 2400 2700

*

Macrophages

Cell content/ml fluid

Fig.4,5,6. Medicinal water bath significantly attenuated the number of inflammatory cells obtained from BAL fluid samples (n=12/group,

*p<0.05, Student t test) Fig.5. Fig.6.

Fig.7. Medicinal water bath did not influence the concentration of IL-1β (n=12/group, p>0.05)

a b c

SROP 4.1.2.B-10/2/KONV-20/0-0002; SROP 4.2.2/B-10/1-2010-0029; OTKA K-73044; OTKA K81984.