Female ICR mice (5-6 weeks old) were purchased from Changzhou Cavens Laboratory Animal Ltd.
(Changzhou, China). The mice were maintained at 25 ± 2℃ with a 12 h light/dark cycle and fed with a standard rat chow and water ad libitum. All experiments were approved by the Ethical Committee for Animal Experiments of Navy Medical University, and strictly carried out in accordance with the approved guidelines.
Mice were randomly divided into four groups (n = 6 each): (1) control group, (2) 60% H 2 group, (3) asthma group, and (4) asthma + 60% H 2 group. The mice were sensitized with ovalbumin (OVA) + Al(OH) 3 (both from Sigma, St Louis, MO) (100 µg OVA and 3 mg Al(OH) 3 in 300 µl PBS) or PBS (300 µl PBS) by subcutaneous injection on day 1, 8 and 15. This was then followed by a nasal drip of 20 µl OVA (5 µg OVA in 20 µl PBS) or PBS (20 µl PBS) on day 22 and 29. To assess the effects of hydrogen gas on asthma, the mice of 60% H 2 group and asthma + 60% H 2 group were housed in a speci c airtight device producing air mixture including 60% H 2 and 21% O 2 for two weeks (2 hours per day).
(Changzhou, China). The mice were maintained at 25 ± 2℃ with a 12 h light/dark cycle and fed with a standard rat chow and water ad libitum. All experiments were approved by the Ethical Committee for Animal Experiments of Navy Medical University, and strictly carried out in accordance with the approved guidelines.
Mice were randomly divided into four groups (n = 6 each): (1) control group, (2) 60% H 2 group, (3) asthma group, and (4) asthma + 60% H 2 group. The mice were sensitized with ovalbumin (OVA) + Al(OH) 3 (both from Sigma, St Louis, MO) (100 µg OVA and 3 mg Al(OH) 3 in 300 µl PBS) or PBS (300 µl PBS) by subcutaneous injection on day 1, 8 and 15. This was then followed by a nasal drip of 20 µl OVA (5 µg OVA in 20 µl PBS) or PBS (20 µl PBS) on day 22 and 29. To assess the effects of hydrogen gas on asthma, the mice of 60% H 2 group and asthma + 60% H 2 group were housed in a speci c airtight device producing air mixture including 60% H 2 and 21% O 2 for two weeks (2 hours per day).