论文部分内容阅读
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 leukotriene receptor 1 (CysLT1) in bronchial epithelial cells and investigate whether IFN-γ is involved in the regulation of leukotriene receptor expression. Methods CysLT1 mRNA of bronchial epithelial cells (BEAS-2B, 16HBE, primary cells) was detected by RT-PCR. The cells were stimulated with IFN-γ for 48 h, CysLT1 mRNA was detected by RT-PCR, and the expression of CysLT1 protein was detected by Western blot. Results The mRNA of CysLT1 was not amplified in bronchial epithelial cells by Taq DNA polymerase RT-PCR, but was replaced by KODTM DNA polymerase in bronchial epithelial cells (BEAS-2B, 16HBE, primary cells) stimulated by IFN-γ Amplified to CysLT1 mRNA. When the IFN-γ levels were 0, 0.1 μg.L-1, 1.0 μg.L-1, 10.0 μg.L-1, 50.0 μg.L-1, and 100.0 μg.L-1 respectively, the CysLT1 mRNA / The relative density ratios of 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. The relative density of CysLT1 protein was 3.09 ± 0.17 and 13.27 ± 0.33 when the IFN-γ levels were 0, 1.0μg.L-1, 10.0μg.L-1, 50.0μg.L-1 and 100.0μg.L-1 respectively , 27.44 ± 0.29, 29.25 ± 0.24, 31.42 ± 0.49 (P <0.01). These results suggest that IFN-γ up-regulates the expression of CysLT1 at mRNA and protein levels in a concentration-dependent manner. Conclusion Bronchial epithelial cells are not only a barrier cell, but also an effector cell that can express leukotriene receptor CysLT1. The large amount of inflammatory mediator IFN-γ produced locally in upper respiratory tract infection upregulates the expression of CysLT1 in epithelial cells and enhances the inflammatory effect of leukotrienes on the respiratory tract, which may be one of the mechanisms of wheezing induced by viral respiratory infection.