地塞米松抑制LPS和TNF-α诱导的人气道平滑肌细胞IL-33表达

发布时间：2018-06-25 04:54

本文选题：人气道平滑肌细胞 + 白介素33　；参考：《华中科技大学》2013年硕士论文

【摘要】：目的：白介素33（IL-33）是最新发现的IL-1家族成员，是一个组织来源的细胞因子，主要位于核内。其受体为ST2，两者结合可以介导Th2免疫反应，而Th2/Th1反应失衡在支气管哮喘的发病中起重要作用。脂多糖（Lipopolysaccharide，LPS）是革兰阴性杆菌的胞壁成分，能介导支气管哮喘炎症反应。LPS通过结合Toll样受体4（TLR-4）参与免疫反应。近年的研究发现气道平滑肌细胞（ASMC）同样能够参与炎症反应，且细胞表面可以表达TLR-4。这篇研究旨在对LPS能否刺激离体培养的人ASMC产生IL-33，作为难治性哮喘可能相关因子之一——TNF-α是否与LPS具有协同刺激作用以及糖皮质激素对IL-33生成的影响展开讨论。方法：手术室收取因肺部疾病行肺段或肺叶切除的人支气管标本（直径0.5-1.0cm）,原代培养支气管平滑肌细胞，选取6-8代平滑肌细胞进行实验。以每孔105cells/cm2种于六孔板中，分别或联合加入各刺激物LPS、TNF-α、哮喘致敏血清（AAS）、IgE。抑制实验则将DXM、SB203580、PD98059等抑制物提前各刺激物30分钟加入。相应时间点收集细胞上清行ELISA，收集细胞提取RNA行实时荧光定量PCR检测。结果： LPS引起人气道平滑肌细胞IL-33基因表达增高（p0.05）;哮喘致敏血清可能促进气道平滑肌细胞IL-33产生，但这种效应与IgE作用关系不大（p0.05）;DXM抑制经LPS和TNF-α诱导的IL-33mRNA生成（p0.05）；SB203580和PD98059部分抑制LPS和TNF-α诱导的IL-33生成。IL-33在细胞上清中的水平实验组与对照组基本一致。结论：糖皮质激素能抑制多种细胞因子的生成而发挥抗炎作用。本实验证明地塞米松能够在体外抑制LPS和TNF-α诱导的IL-33基因表达，而p38MAPK和ERK通路抑制剂不能完全阻断这种作用。目前糖皮质激素在哮喘控制和治疗中仍然有重要地位。
[Abstract]:Aim: interleukin-33 (IL-33) is a newly discovered member of the IL-1 family. It is a tissue-derived cytokine mainly located in the nucleus. The receptor is ST2, which can mediate Th2 immune response, and the imbalance of Th2 / Th1 response plays an important role in the pathogenesis of bronchial asthma. Lipopolysaccharide (LPS) is a cell wall component of Gram-negative bacilli, which mediates the inflammatory response of bronchial asthma. LPS takes part in the immune response by binding Toll-like receptor 4 (TLR-4). In recent years, it has been found that airway smooth muscle cells (ASMC) can also participate in inflammatory response, and the surface of the cells can express TLR-4. The aim of this study was to investigate whether LPS could stimulate the production of IL-33 in cultured human ASMC, and whether TNF- 伪 could co-stimulate with LPS and the effect of glucocorticoid on IL-33 production as one of the possible factors associated with refractory asthma. Methods: the primary cultured bronchial smooth muscle cells (BSMCs) were collected from human bronchial specimens (diameter 0.5-1.0cm) after lung segmenting or lobectomy in the operating room, and 6-8 passages of smooth muscle cells were selected for the experiment. 105cells/cm2 was seeded into the six-hole plate with the addition of each stimulator LPS-TNF- 伪, and the asthma sensitized serum (AAS) IgE. In the inhibition experiment, DXMN SB203580, PD98059 and other inhibitors were added 30 minutes in advance. ELISAs were collected from the supernatant of the cells collected at the corresponding time points, and RNA extracted from the collected cells was detected by real-time fluorescence quantitative polymerase chain reaction (FQ-PCR). Results: LPS-induced increase of IL-33 gene expression in human airway smooth muscle cells (p0.05), asthma sensitized serum may promote IL-33 production in airway smooth muscle cells, but this effect is not related to the effect of IgE (p0.05) DXM inhibits IL-33 mRNA production induced by LPS and TNF- 伪 (p0.05). SB203580 and PD98059 partially inhibited the production of IL-33 induced by LPS and TNF- 伪. The level of IL-33 in the supernatant of the experimental group was similar to that of the control group. Conclusion: glucocorticoid can inhibit the production of many cytokines and play an anti-inflammatory effect. This study demonstrated that dexamethasone could inhibit the expression of IL-33 gene induced by LPS and TNF- 伪 in vitro, but p38 MAPK and ERK pathway inhibitors could not completely block this effect. At present, glucocorticoids still play an important role in asthma control and treatment.
【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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