哮喘小鼠肺组织和肺T细胞β1,3-N-乙酰糖基转移酶的表达

Expression of Fringe in the Asthmatic Mouse Model

目的研究支气管哮喘模型小鼠肺组织及肺T细胞中三种β1,3-N-乙酰糖基转移酶(Fringe)RFNG、LFNG、MFNG的基因和蛋白表达的特点,探讨Fringe是否与哮喘发病有关。方法使用卵白蛋白腹腔注射后雾化吸入法制备BALB/c小鼠哮喘模型,并设置生理盐水对照组。经Perco1及尼龙毛法分离获取肺T细胞。采用半定量RT-PCR检测两组小鼠肺组织及肺T细胞中RFNG、LFNG、MFNGmRNA表达;Westernblot检测两组小鼠肺组织中RFNG、LFNG、MFNG蛋白表达。结果两组小鼠肺组织及肺T细胞中均存在三种Fringe表达。哮喘组肺组织及肺T细胞的RFNGmRNA表达较对照组增加(肺组织:0.92±0.35比0.51±0.13,P<0.01;肺T细胞:0.33±0.06比0.18±0.07,P<0.01);LFNGmRNA表达较对照组减少(肺组织:0.77±0.32比1.61±0.31,P<0.01;肺T细胞:0.49±0.19比0.71±0.03,P<0.01)。MFNG在肺组织中的表达无明显差异(肺组织:1.44±0.29比1.70±0.44,P>0.05),但MFNG在肺T细胞中的表达明显减少(0.11±0.03比0.52±0.18,P<0.01)。肺组织中三种Fringe蛋白表达与肺组织的mRNA结果相似。哮喘组的RFNG蛋白表达高于对照组(1.17±0.04比0.68±0.07,P<0.05),MFNG蛋白表达在两者之间没有明显的差异(8.10±0.60比9.12±0.07,P>0.05),而LFNG的蛋白表达则低于对照组(4.11±0.38比6.41±0.11,P<0.05)。结论三种Fringe的异常表达可能与支气管哮喘的发病有关。

Objective To investigate the expressions of β1,3-N-acetyl glucosaminyl transfrases（Fringe）（RFNG,LFNG and MFNG）in lung tissues and lung T cells isolated from asthmatic mice,and to explore the role of Fringe in pathogenesis of asthma.Methods Asthmatic BALB/c mouse model was established by inhalation of ovalbumin after intraperitoneal injection.Meanwhile,the control groups were established by normal saline.Lung tissues were sampled after 24 hours since the last stimulation.T cells were isolated from the lung tissues using percol and NylonFiber.The mRNA expressions of three kinds of Fringe in the lung tissues and lung T cells were examined by reverse transcription-PCR（RT-PCR）.The protein expressions of Fringe in the lung tissues were detected by Western blot.Results The mRNA expressions of RFNG,LFNG and MFNG were detectable in the lung tissues and lung T cells.The mRNA expressions of RFNG increased in the asthmatic group compared with the control group（lung tissues：0.92±0.35 vs 0.51±0.13,P0.01;lung T cells：0.33±0.06 vs 0.18±0.07,P0.01）.LFNG mRNA had lower expression level in the asthmatic group（lung tissue：0.77±0.32 vs 1.61±0.31,P0.01;lung T cells：0.49±0.19 vs 0.71±0.03,P0.01）.No difference on the mRNA expression of MFNG was found in the lung tissues（1.44±0.29 vs 1.70±0.44,P0.05）.MFNG mRNA expression decreased in the asthmatic group compared with the control group in the T cells（1.17±0.04 vs 0.68±0.07,P0.05）.The results of western blot were consistent with RT-PCR results of the lung tissues.The expressions of RFNG increased in the asthmatic group（1.17±0.04 vs 0.68±0.07,P0.05）.The expression of MFNG has no difference between two groups（8.10±0.60 vs 9.12±0.07,P0.05）.LFNG had a lower expression in the asthmatic group（4.11±0.38 vs 6.41±0.11,P0.05）.Conclusion The abnormal expressions of three kinds of Fringe may be involved in the pathogenesis of asthma.