IL-4和IL-13对支气管上皮细胞上CXC趋化因子受体CXCR4表达的影响

发布时间：2018-04-21 13:43

本文选题：IL-4 + IL-13　；参考：《华中科技大学》2013年硕士论文

【摘要】：研究背景： CXCR4存在于机体多种细胞表面，是趋化因子SDF-1α的特异性受体，在细胞的增殖和迁移中起着重要作用。有研究证实CXCR4及其配体SDF-1α参与了哮喘的气道炎症和气道高反应，在哮喘中发挥重要作用。Th2细胞因子IL-4和IL-13在哮喘的发病机制过程中扮演了重要角色，嗜酸性粒细胞表面CXCR4的表达在IL-4和IL-13的影响下有所改变。而哮喘中支气管上皮细胞是否表达CXCR4以及其表达是否受Th2细胞因子IL-4和IL-13的影响，，目前尚未有研究报道。目的：探讨支气管上皮细胞在Th2细胞因子环境下是否表达CXCR4以及其表达是否受IL-4和IL-13的影响。方法：本实验通过用六孔板培养人支气管上皮细胞系，饥饿16h，然后用IL-4（20ng/ml）和IL-13（20ng/ml）分别及联合刺激支气管上皮细胞24h，形成Th2细胞因子环境，以体外模拟哮喘环境。通过提取蛋白，电泳，把丙烯酰胺凝胶中的蛋白转移到PVDF膜上，封闭1h，CXCR4单克隆抗体抗体孵育过夜，洗涤后加入二抗孵育1h，洗涤，ECL显色、曝光。结果：正常组HBEsCXCR4蛋白表达灰度比值为（0.54±0.07），而IL-4刺激组CXCR4蛋白表达灰度比值为（0.91±0.07），明显高于正常组（P0.05），具有统计学差异。IL-13刺激组CXCR4蛋白表达灰度比值为（1.11±0.11），与正常组比较，也明显增高（P0.05），差异具有统计学意义。IL-4+IL-13联合刺激组CXCR4蛋白表达灰度比值（1.19±0.18），明显高于正常组（P0.05），具有统计学差异。结论：该实验结果表明哮喘环境中支气管上皮细胞表面CXCR4蛋白表达上调，Th2细胞因子IL-4和IL-13均可使支气管上皮细胞表面CXCR4表达增高，说明其表达受到Th2细胞因子的调控。
[Abstract]:Background: CXCR4 is a specific receptor of chemokine SDF-1 伪 and plays an important role in cell proliferation and migration. It has been confirmed that CXCR4 and its ligand SDF-1 伪 are involved in airway inflammation and airway hyperresponsiveness in asthma. Th2 cytokines, IL-4 and IL-13, play an important role in the pathogenesis of asthma. The expression of CXCR4 on eosinophil surface was affected by IL-4 and IL-13. However, whether bronchial epithelial cells express CXCR4 and whether their expression is affected by Th2 cytokines, IL-4 and IL-13, has not been reported. Objective: To investigate whether bronchial epithelial cells express CXCR4 in the environment of Th2 cytokines and whether their expression is affected by IL-4 and IL-13. Methods: In this experiment, human bronchial epithelial cell line was cultured with six-hole plate for 16 h, then stimulated with IL-4 + 20 ng / ml and IL-13 + 20 ng / ml for 24 h, respectively, to form Th2 cytokine environment, and to simulate asthma in vitro. The protein in acrylamide gel was transferred to PVDF membrane by extracting protein and electrophoretic. The monoclonal antibody against CXCR4 was incubated overnight after washing, then the second antibody was added for 1 hour. Results: The gray level ratio of HBEsCXCR4 protein expression in normal group was 0.54 卤0.07, while that of CXCR4 protein in IL-4 stimulation group was 0.91 卤0.07, which was significantly higher than that in normal group (P 0.05). The ratio of CXCR4 protein expression in IL-13 stimulated group was 1.11 卤0.11, which was significantly higher than that in normal group. The grayscale ratio of CXCR4 protein expression in IL-4 IL-13 combined stimulation group was 1.19 卤0.18, which was significantly higher than that in normal control group (P 0.05). Conclusion: The results showed that the expression of Th _ 2 cytokine IL-4 and IL-13 could increase the expression of CXCR4 on the surface of bronchial epithelial cells in asthmatic environment, which indicated that the expression of Th2 cytokines regulated the expression of Th _ 2 cytokines.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R562.25

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