摘要
目的评价虫草素及虫草素与糖皮质激素联合应用对慢性哮喘模型小鼠气道平滑肌收缩的调节作用及可能机制。方法将32只C57／BL6小鼠随机分成对照组和慢性哮喘模型组，利用肌动描记仪比较p38蛋白激酶抑制剂SB239063（10^-6M）、虫草素（10^-6M）以及糖皮质激素地塞米松（10^-6M）单独或联合孵育前、后由乙酰胆碱（acetyleholine，Ach）诱导的支气管收缩反应，同时应用蛋白印迹检测支气管组织中p38蛋白激酶和热休克蛋白27（heatshockproteins27，HSP27）的磷酸化状态。结果模型组小鼠的支气管收缩较对照组加重，有统计学意义。对照组和模型组p38蛋白激酶抑制剂SB239063孵育1h后对Ach介导的收缩反应较空白对照组下降（P〈0．05），地塞米松单独和地塞米松联合虫草素在模型组小鼠Ach诱导支气管平滑肌收缩力的作用均较对照组降低（P〈0．05）；地塞米松单独和地塞米松联合虫草素孵育后，对照组和模型组小鼠支气管组织p38蛋白激酶和HSP27磷酸化水平较Ach单纯作用组降低，而且在模型组，使用虫草素干预后p38蛋白激酶和HSP27磷酸化水平较Ach单纯作用组下降（P值均〈0．05）。结论Ach诱导的离体支气管收缩反应具有良好的可重复性，卵清白蛋白吸入可导致Ach诱导的支气管收缩反应加重，糖皮质激素联合虫草素治疗较糖皮质激素单独治疗更有效抑制Ach诱导的气道平滑肌收缩，此种潜在的协同作用可能通过更大程度地抑制p38蛋白激酶信号通路为机制。
Objective To investigate the role of cordycepin on airway smooth muscle contraction and the potential role of p38 mitogen-activated protein kinases （p38MAPK） in airway smooth muscle contractile response. Methods C57/BL6 mice were divided into air-exposure and ovalbumin （OVA） exposure group, the bronchi were isolated from them and incubated with vehicle, p38MAPK inhibitor SB239063 （ 10.6 M）, Dexamethasone （Dex） and/or codycepin （ 10.6 M）. Acetylcholine （Ach） induced bronchial contractile responses were measured before and after incubation using myograph. Western blot analysis was used to measure the phosphorylation of p38MAPK and heat shock proteins 27（HSP27）. Results Bronchial contractile responses to Ach were significantly enhanced in OVA exposed mice than in air exposed mice. Either 10 6 M SB239063 or 10s M Dex and/or 10-6 M cordycepin could equally inhibit Ach induced bronchial contraction Emax in both groups （ P 〈0.05）, while cordycepin alone failed to do so. Dex and/or cordycepin could inhibit the phosphorylation of p38MAPK than Ach stimulated group significantly in air/OVA exposure mice, while the remarkably inhibition was shown in OVA exposure mice when pretreated with cordycepin,Dex and/or cordycepin could inhibit the phosphorylation of HSP27 than Ach stimulated group significantly in air/OVA exposure mice. Conclusions Ach induced exvivo bronchial contractility was highly reproducible, while OVA exposure could enhance the airway smootl muscle contraction, p38MAPK was involved in Ach induced airway smooth muscle contractility. Dex and Cordyeepin could more effectively inhibit airway smooth muscle contraction than Dex alone, those effects may he p38MAPK/HSP27 dependent.
出处
《国际呼吸杂志》
2012年第20期1545-1551,共7页
International Journal of Respiration
基金
国家自然基金项目（81070023）,国家“十一五”863计划课题资助项目
