Lyn对屋尘螨诱导的人支气管上皮细胞中MUC5AC表达的影响及其机制

目的：探讨Lyn对屋尘螨（HDM）诱导的人支气管上皮细胞中MUC5AC表达的影响，并初步阐明其可能的相关作用机制。方法：选取人支气管上皮16HBE细胞，将其分为PBS组（给予PBS）和HDM组（给予1 μg·L^-1HDM），采用脂质体转染法将LynsiRNA分别转至PBS组和HDM组细胞。构建人MUC5AC启动子荧光素酶报告基因，采用Promega双荧光素酶报告基因试剂盒检测相对荧光素酶活性（RLU），采用免疫荧光法检测16HBE细胞中MUC5AC的表达，并在共聚焦显微镜下观察。采用Western blotting法检测16HBE细胞中STAT6的表达水平。结果：双荧光素酶报告基因检测，HDM组16HBE细胞中MUC5AC启动子的RLU高于PBS组（P<0.05）；与未干扰时比较，LynsiRNA干扰后HDM组RLU明显升高（P<0.05）。免疫荧光法检测，HDM组MUC5AC表达水平高于PBS组（P<0.05）；与未干扰时比较，LynsiRNA干扰后HDM组MUC5AC表达水平升高（P<0.05）。Western blotting法检测，LynsiRNA干扰后，HDM组细胞中STAT6表达水平升高（P<0.05）。结论：Lyn缺失能增加人支气管上皮细胞中MUC5AC的表达，其机制可能与Lyn调控STAT6信号途径有关。

Effect of Lyn on expression of MUC5AC in human bronchial epithelial cells induced by house dust mite and its mechanism

Objective:To investigate the effect of Lyn on the expression of MUC5AC in human bronchial epithelial cells induced by house dust mite(HDM),and to explore its possible mechanism.Methods:The human bronchial epithelial cells(16 HBE)were divided into PBS group and HDM group(1 μg·L-1HDM).The cells were transfected by liposome.The luciferase report gene of MUC5AC promoter was constructed.The relative luciferase unit(RLU)was detected by double luciferase report gene assay.The expression of MUC5AC in the cells was detected by immunofluorescence technique and observed by confocal microscope.The expression level of STAT6 in the 16HBE cells was detected by Western blotting method.Results:The results of double luciferase report gene assay showed that the RLU in HDM group was higher than that in PBS group(P<0.05).The RLU of cells in HDM group treated with LynsiRNA intervention was higher than that of the cells without LynsiRNA intervention(P<0.05).The immunofluorescence results demonstrated that the expression level of MUC5AC in HDM group was higher than that in PBS group(P<0.05),and the expression level of MUC5AC in HDM group was increased after LynsiRNA intervention(P<0.05).The Western blotting results indicated that the expression level of STAT6 was up-regulated in HDM group when the cells were intervened with LynsiRNA(P<0.05). Conclusion:Deficiencyof Lyn can increase the expression of MUC5AC in the human bronchial epithelial cells,and its mechanism may be related to regulating the STAT6 signal pathway by Lyn.