Effect of intranasal T-bet plasmid transfer on bronchial alveolar lavage fluid IL-13 and ET-1 in a murine asthmatic model

Abstract

Objective To investigate the effects of airway T-bet plasmid gene transfer on IL-13 and ET-1 in bronchial alveolar lavage fluid (BALF) of murine asthmatic model. Methods Thirty-two C57BL/6J mice were randomized into four groups (8 mice each):the asthmatic model group (group A) , the normal control group (group B) , the pcDNA3 plasmid group (group C) and the pcDNA3-T-bet group (group D). The mouse asthma model was established by sensitization with intraperitoneal injection of ovalbumin (OVA) and intranasal challenge in all animals except for the group B where normal saline was used instead. At 48 hours before intranasal challenges, the group C was intranasally administered 50 μg pcDNA3 plasmid, and the group D with 50μg pcDNA3-T-bet plasmid.At 48 hours after OVA challenge, BALF was collected for determination of IL-13 and ET-1 levels by immunoblotting and ELISA. Results The levels of IL-13 and ET1 in BALF were significantly elevated in the asthmatic model group compared with the normal control group [(2.32±0.40)ng/L vs (0.38±0.16)ng/L,(40.07±6.52)ng/L vs (l4.16±0.5l)ng/L, both P＜0.05), and were lowered in the pcDNA3-T-bet group[(1.16±0.19)ng/L and (23.08±2.59)ng/L, respectively] compared with the asthmatic model group (both P＜0.05). Conclusion Intranasal pcDNA3-T-bet plasmid transfer can reduce the levels of IL-13 and ET-1 in BALF of the murine asthmatic model. Key words: Bronchoalveolar lavage fluid; Asthma; T - bet genes; Interleukin 13; Endothelin 1