南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (3): 347-353.doi: 10.12122/j.issn.1673-4254.2022.03.05

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HSP90α通过调控气道上皮细胞内质网应激加重屋尘螨诱导的哮喘气道炎症

黄浩华，乔妤婕，黄 奕，董航明   

  1. 南方医科大学南方医院呼吸与危重症医学科，广东 广州 510515
  • 出版日期:2022-03-20 发布日期:2022-04-11

HSP90α exacerbates house dust mite-induced asthmatic airway inflammation by upregulating endoplasmic reticulum stress in bronchial epithelial cells

HUANG Haohua, QIAO Yujie, HUANG Yi, DONG Hangming   

  1. Department of Respiratory and Critical Care Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China
  • Online:2022-03-20 Published:2022-04-11

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摘要： 目的 探讨热休克蛋白90（HSP90α）参与屋尘螨（HDM）诱导的哮喘气道炎症的作用及调控气道上皮细胞内质网应激的机制。方法 人气道上皮细胞HBE体外培养，由HDM诱导哮喘体外模型，浓度梯度刺激24 h，分为正常对照组；200 U/mL组；400 U/mL组；800 U/mL组。随后分别使用siHSP90α干扰序列转染及HSP90抑制剂17-AAG干预，分为HBE正常对照组、HBE+HDM组（800 U/mL，24 h）、HBE+HDM（800 U/mL，24 h）+siHSP90α（50 nmol，12 h）组、HBE+HDM（800 U/mL，24 h）+17-AAG（900 nmol，6 h）组，测定HSP90α以及内质网应激标志物的表达水平。C57BL/6小鼠由HDM诱导哮喘体内模型，分为正常对照组、17-AAG滴鼻组、HDM滴鼻组、HDM+17-AAG滴鼻组，测定气道炎症水平与内质网应激标志物的表达水平。结果 在HDM诱导的HBE细胞哮喘体外模型中，蛋白电泳结果显示HSP90α（P=0.011）表达上调，琼脂糖凝胶电泳结果显示内质网应激标志物XBP-1（P=0.044）、ATF-6α（P=0.030）和GRP-78（P=0.027）表达水平显著上调；而敲低HSP90α和使用17-AAG会显著抑制HDM诱导的XBP-1（P=0.008）上调；在HDM诱导的哮喘小鼠模型中，相较于HDM组，H&E染色和瑞氏-姬萨姆染色显示HDM+17-AAG组的气道炎症改善，炎症细胞聚集明显减少。肺泡灌洗液计数显示，炎症细胞数量显著减少（P=0.014）。ELISA法显示肺泡灌洗液中炎症因子IL-4（P=0.030）、IL-5（P=0.035）显著降低。免疫组织化学染色显示气道上皮细胞XBP-1和GRP-78表达明显减少。结论 HSP90α加重HDM诱导的哮喘气道炎症，可能通过上调内质网应激实现的。

关键词: 哮喘；内质网应激；HSP90α；屋尘螨

Abstract: Objective To explore the role of heat shock protein 90α (HSP90α) and endoplasmic reticulum (ER) stress pathway in allergic airway inflammation induced by house dust mite (HDM) in bronchial epithelial cells. Methods A HDM- induced asthmatic cell model was established in human bronchial epithelial (HBE) cells by exposure to a concentration gradient (200, 400 and 800 U/mL) of HDM for 24 h. To test the effect of siHSP90α and HSP90 inhibitor 17-AAG on HDM-induced asthmatic inflammation, HBE cells were transfected with siHSP90α (50 nmol, 12 h) or pretreated with 17-AAG (900 nmol, 6 h) prior to HDM exposure (800 U/mL) for 24 h, and the changes in the expression of HSP90α and ER stress markers were assessed. We also tested the effect of nasal drip of 17-AAG, HDM, or their combination on airway inflammation and ER stress in C57BL/6 mice. Results In HBE cells, HDM exposure significantly up-regulated the expression of HSP90α protein (P=0.011) and ER stress markers XBP-1 (P=0.044), ATF-6α (P=0.030) and GRP-78 (P=0.027). Knocking down HSP90α and treatment with 17-AAG both significantly inhibited HDM-induced upregulation of XBP-1 (P=0.008). In C57BL/6 mice, treatment with 17-AAG obviously improved HDM-induced airway inflammation and significantly reduced the number of inflammatory cells in the airway (P=0.014) and lowered the levels of IL-4 (P=0.030) and IL-5 (P=0.035) in alveolar lavage fluid. Immunohistochemical staining showed that the expressions of XBP-1 and GRP-78 in airway epithelial cells decreased significantly after the treatment of 17-AAG. Conclusions HSP90α promotes HDM-induced airway allergic inflammation possibly by upregulating ER stress pathway in bronchial epithelial cells.

Key words: asthma; endoplasmic reticulum stress; heat shock protein 90α; house dust mite