SB203580对糖皮质激素敏感性的影响及作用机制的研究

发布时间：2018-04-05 04:14

  本文选题：支气管哮喘　切入点：糖皮质激素　出处：《北京协和医学院》2012年博士论文

【摘要】：目的建立烟雾暴露的哮喘大鼠模型及尼古丁诱导人肺腺癌A549细胞糖皮质激素抵抗的实验模型,观察p38丝裂原活化蛋白激酶(p38MAPK)抑制剂SB203580对烟雾暴露的哮喘大鼠的影响,并分别从体内实验、体外实验两个方面探讨SB203580影响糖皮质激素敏感性的机制。 方法Wistar大鼠随机分为4组,即正常对照组、哮喘组、烟雾暴露的哮喘组及SB203580干预组。动物肺功能仪测定大鼠呼气阻力、吸气阻力及肺顺应性,观察肺组织病理学改变,通过ELISA检测大鼠肺组织匀浆中IL-4、IL-5和IL-8的水平。经RT-PCR检测大鼠肺组织糖皮质激素受体(GR)、HSP90和p38MAPK mRNA的表达,经Western blot检测大鼠肺组织GR、HSP90和p38MAPK蛋白的表达。培养A549细胞,将其随机分为四组：A组：空白对照组,B组：1umol/L的地塞米松,C组：1umol/L的地塞米松+1umol/L的尼古丁,D组：1umol/L的地塞米松+1umol/L的尼古丁+1umol/L的SB203580,采用免疫荧光染色技术分析GR亚细胞定位。 结果与烟雾暴露的哮喘组相比,SB203580干预组大鼠的气道阻力显著下降,肺顺应性显著升高,差异有统计学意义(P0.05)；气道炎症明显减轻；肺组织中IL-4、IL-5和IL-8的含量显著下降,差异有统计学意义(P0.05)；肺组织GRmRNA的表达无明显变化,HSP90和p38MAPK mRNA的表达显著下降,差异有统计学意义(P0.05)；肺组织GR、HSP90和p38MAPK蛋白表达呈现与mRNA表达相一致的结果。C组A549细胞GR的核浆比为0.08±0.05,D组A549细胞GR的核浆比为0.59±0.25,两组之间的差异有统计学意义(P0.05)。 结论p38MAPK抑制剂SB203580可以改善烟雾暴露的哮喘大鼠的气道炎症,减轻其支气管收缩反应。其通过促使GR核转位提高糖皮质激素敏感性。
[Abstract]:Objective to establish a rat model of asthma induced by smoke exposure and an experimental model of nicotine induced glucocorticoid resistance in human lung adenocarcinoma cell line A549, and to observe the effect of p38 mitogen activated protein kinase (p38 MAPK) inhibitor SB203580 on smog exposed asthmatic rats.The effects of SB203580 on glucocorticoid sensitivity were investigated in vivo and in vitro.Methods Wistar rats were randomly divided into 4 groups: normal control group, asthma group, smoke exposed asthma group and SB203580 intervention group.The expiratory resistance, inspiratory resistance and lung compliance in rats were measured by animal pulmonary function analyzer. The histopathological changes of lung tissue were observed. The levels of IL-4 IL-5 and IL-8 in rat lung homogenate were detected by ELISA.The expression of glucocorticoid receptor (GRN) HSP90 and p38MAPK mRNA in rat lung tissue was detected by RT-PCR, and the expression of GRN HSP90 and p38MAPK protein in rat lung tissue was detected by Western blot.A549 cells were cultured,灏嗗叾闅忔満鍒嗕负鍥涚粍:A缁，

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