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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

摘要:

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.

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abstracts:

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.

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作者: 徐小彩 [1] 檀卫平 [1] 吴葆菁 [1] 麦贤弟 [1] 黄花荣 [1]
作者单位: 中山大学附属第二医院儿科,广州,510120 [1]
分类号: R562.25
栏目名称: 论著
DOI: 10.3760/cma.j.issn.1674-1927.2011.02.005
发布时间: 2011-09-08
基金项目:
广东省自然科学基金 广东省科委科技计划项目 2009年度广东省自然科学基金资助博士科研启动项目
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