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目的观察单免疫球蛋白白细胞介素1受体相关蛋白(SIGIRR)对香烟烟雾提取物(CSE)刺激的小鼠肺泡上皮细胞株A549炎性反应的影响。方法将A549细胞分为过表达SIGIRR组和对照组。构建含有SIGIRR c DNA全长的真核表达载体pc DNA3.1(+),瞬时转染小鼠Ⅱ型肺泡上皮细胞来源的A549细胞,使SIGIRR在A549细胞中过表达,采用半定量Western blot和RT-PCR法检测其表达。以CSE刺激对照组及过表达SIGIRR组中A549细胞后,ELISA法检测白细胞介素6(IL-6)水平,双荧光素酶报告系统检测环氧化酶2(COX-2)表达水平,化学发光法检测活性氧(ROS)释放水平的变化。结果转染SIGIRR表达载体的A549细胞其SIGIRR表达量显著上升。对照组经CSE刺激后,COX-2表达、ROS释放、IL-6释放水平均显著增高,而过表达SIGIRR组的A549细胞经CSE刺激后,COX-2表达较对照组降低,ROS释放较对照组降低,IL-6释放水平也明显低于对照组(P<0.05)。结论 SIGIRR表达上调可以抑制CSE诱导的A549细胞产生ROS、COX-2及IL-6,提示SIGIRR表达上调可能对吸烟所致呼吸道炎症具有抑制作用。
Objective To investigate the effects of single immunoglobulin interleukin 1 receptor related protein (SIGIRR) on the inflammatory response of mouse alveolar epithelial cell line A549 stimulated by cigarette smoke extract (CSE). Methods A549 cells were divided into overexpression SIGIRR group and control group. The full-length eukaryotic expression vector pcDNA3.1 (+) containing SIGIRR c DNA was constructed and transiently transfected into A549 cells of mouse type II alveolar epithelial cells. SIGIRR was overexpressed in A549 cells. RT-PCR method to detect the expression. A549 cells were stimulated with CSE and SIGIRR cells were overexpressed. The levels of IL-6 were detected by ELISA. The expression of cyclooxygenase 2 (COX-2) was detected by dual luciferase reporter assay. Luminescence assay was used to detect changes of reactive oxygen species (ROS) release. Results The expression of SIGIRR in A549 cells transfected with SIGIRR expression vector was significantly increased. Compared with the control group, COX-2 expression, ROS release and IL-6 release were significantly increased in the control group after CSE stimulation, while COX-2 expression in A549 cells overexpressing SIGIRR group was lower than that in the control group Group decreased, IL-6 release was also significantly lower than the control group (P <0.05). Conclusion Upregulation of SIGIRR can inhibit the production of ROS, COX-2 and IL-6 in A549 cells induced by CSE, suggesting that the upregulation of SIGIRR may inhibit the respiratory inflammation induced by smoking.