干扰素-γ对支气管上皮细胞表达白三烯受体半胱氨酰白三烯受体1的调节  

Modulation of Leukotriene Receptor Cysteinyl Leukotriene Recepter 1 Expression in Bronchial Epithelial Cells by Interferon-γ

作　　者：欧维琳[1] 朱春江[1] 蒋林彬[1] 韦欢[1] 张明[1] 宋晓丹[1] 吉艳荣[1] 

机构地区：[1]桂林医学院附属医院儿科,广西桂林541001

出　　处：《实用儿科临床杂志》2010年第21期1619-1621,1624,共4页Journal of Applied Clinical Pediatrics

基　　金：广西科技厅自然科学基金(桂科0728228)

摘　　要：目的研究白三烯受体半胱氨酰白三烯受体1(CysLT1)在支气管上皮细胞中的表达,探讨炎性因子IFN-γ是否参与调节白三烯受体的表达。方法反转录-PCR(RT-PCR)法检测支气管上皮细胞(BEAS-2B、16HBE、原代细胞)CysLT1 mRNA。采用IFN-γ刺激细胞48 h,RT-PCR法检测CysLT1 mRNA,Western blot法检测细胞膜CysLT1蛋白表达。结果采用Taq DNA聚合酶RT-PCR法在支气管上皮细胞上未能扩增到CysLT1 mRNA,改用KODTMDNA聚合酶在经过IFN-γ刺激的支气管上皮细胞(BEAS-2B、16HBE、原代细胞)中扩增到CysLT1 mRNA。当IFN-γ水平分别为0、0.1μg.L-1、1.0μg.L-1、10.0μg.L-1、50.0μg.L-1、100.0μg.L-1时,CysLT1 mRNA/β-actin相对密度比为0.420±0.013、0.560±0.008、0.680±0.011、1.230±0.023、1.350±0.016、1.880±0.021。当IFN-γ水平分别为0、1.0μg.L-1、10.0μg.L-1、50.0μg.L-1、100.0μg.L-1时,CysLT1蛋白相对密度为3.09±0.17、13.27±0.33、27.44±0.29、29.25±0.24、31.42±0.49(P<0.01)。提示IFN-γ能上调CysLT1在mRNA水平和蛋白水平的表达,这种上调作用呈浓度依赖性。结论支气管上皮细胞不仅是一种屏障细胞,也是一种效应细胞,能表达炎性介质白三烯受体CysLT1。上呼吸道感染时局部产生的大量炎性介质IFN-γ,使上皮细胞CysLT1表达上调,增强了白三烯对呼吸道的炎症效应,这可能是病毒性呼吸道感染诱发喘息的机制之一。Objective To investigate the expression of leukotriene receptor cysteinyl leukotrienes 1 recepter（CysLT1） in bronchial epithelial cells and to explore whether interferon-γ（IFN-γ） involving the modulation of CysLT1 expression.Methods CysLT1 mRNA in bronchial epithelial cells（BEAS-2B,16HBE,primary bronchial epithelial cells） was detected by RT-PCR.BEAS-2B cells stimulated with IFN-γ for 48 hours,CysLT1 mRNA was measured by RT-PCR.CysLT1 in the cell membrane was checked by Western blot.Results CysLT1 mRNA could not be amplified by conventional Taq DNA polymerase from bronchial epithelial cells.CysLT1 mRNA expression was observed by KODTM DNA polymerase in human airway epithelial cells（BEAS-2B,16HBE,and primary bronchial epithelial cells）treated by IFN-γ.When the concentrations of IFN-γ were repectively 0,0.1 μg·L^-1,1.0 μg·L^-1,10.0 μg·L^-1,50.0 μg·L^-1,100.0 μg·L^-1,the relative intensity of CysLT1 mRNA /β-actin were 0.420±0.013,0.560±0.008,0.680±0.011,1.230±0.023,1.350±0.016,1.880±0.021.When the concentrations of IFN-γ were repectively 0,1.0 μg·L^-1,10.0 μg·L^-1,50.0 μg·L^-1,100.0 μg·L^-1,the intensity of CysLT1 protein expression were 3.09±0.17,13.27±0.33,27.44±0.29,29.25±0.24,31.42±0.49（P0.01）.IFN-γ upregulated CysLT1 mRNA and protein expression in cells above with a dose-dependent manner.Conclusions Bronchial epithelial cells are not only barrier but also effect cells.It can express leukotrienes receptor CysLT1,the inflammatory infactor.When upper respiratory tract infected by virus,it produces abundant IFN-γ.Then it upregulated the expression of CysLT1 mRNA and proteine from epithelial cells,enhanced inflammational reaction in respiratory tract.It may explain the mechanism partially that virus respiratory tract infection induced exacerbation of asthma.

关 键 词：白三烯受体 半胱氨酰白三烯 表达 调节 支气管上皮细胞 

分 类 号：R725.6[医药卫生—儿科]