T—bet质粒基因转染对哮喘小鼠支气管肺泡灌洗液白细胞介素13和内皮素1的影响

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

目的 观察气道内T-bet质粒基因转染对哮喘小鼠气道肺泡灌洗液白细胞介素13（IL13）、内皮素1（ET-1）的影响.方法 C57BL/6J小鼠32只,完全随机分为4组（n=8）,即哮喘模型组（A组）、正常对照组（B组）、空质粒干预组（C组）和模型T-bet质粒干预组（D组）.用卵白蛋白（OVA）抗原溶液小鼠腹腔注射致敏,滴鼻造模.B组用生理盐水代替卵白蛋白,C组和D组卵白蛋白激发48 h前,分别经鼻滴入50μg空质粒或重组T-bet质粒.免疫印迹检测和酶联免疫吸附法（ELISA）检测各组小鼠的支气管肺泡灌洗液中IL-13、ET-1的水平.结果 哮喘模型组小鼠支气管肺泡灌洗液（BALF）中Th2因子IL-13和ET-1水平比正常对照组升高[（2.32±0.40）ng/L比（0.38±0.16）ng/L,（40.07±6.52）ng/L比（14.16±0.51）ng/L,均P＜0.05];模型T-bet质粒干预组BALF中Th2因子IL-13和ET-1水平[（1.16±0.19）和（23.08±2.59）ng/L]比哮喘模型组降低（均P＜0.05）.结论 气道内转染T-bet质粒能有效下调小鼠BALF中Th2因子IL-13及ET-1水平.

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.