南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (8): 1333-1338.doi: 10.12122/j.issn.1673-4254.2023.08.09

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VDAC1通过诱导气道上皮细胞铁死亡参与屋尘螨诱导的哮喘小鼠气道炎症

黄 奕，林丽珊，黄浩华，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:2023-08-20 发布日期:2023-09-13

VDAC1 participates in house dust mite- induced asthmatic airway inflammation in mice by inducing ferroptosis of airway epithelial cells

HUANG Yi, Lin Lishan, HUANG Haohua, DONG Hangming   

  1. Department of Respiratory and Critical Care Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China
  • Online:2023-08-20 Published:2023-09-13

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摘要： 目的 探讨电压依赖性阴离子选择性通道蛋白1（VDAC1）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铁死亡的机制。方法 人气道上皮细胞（HBE）体外培养，由HDM诱导HBE细胞进行体外实验，浓度梯度刺激24 h，分为正常对照组；200 U刺激组；400U刺激组；800U刺激组。随后使用VDAC1抑制剂VBIT-4干预，分为HBE正常对照组、VBIT-4组（10 μmol/L，24 h）、HDM（800 U/mL，24 h）组、HDM（800 U/mL，24 h）+VBIT-4（10 μmol/L，24 h）组，测定VDAC1以及铁死亡标志物蛋白的表达水平。使用BALB/c小鼠给予HDM构建哮喘小鼠模型，分为正常对照组、VBIT-4滴鼻组、HDM滴鼻组、HDM+VBIT-4滴鼻组，测定气道炎症水平与铁死亡蛋白的表达水平。结果 通过使用HDM诱导的HBE细胞后，MtROS生成增多，线粒体膜电位下降，免疫印迹试验结果显示VDAC1（P=0.005）表达上调，铁死亡标记物标志物GPX4（P=0.015）表达水平显著降低，FTH1（P=0.030）水平则出现上调；而使用VBIT-4会显著抑制因HDM诱导的FTH1（P=0.037）的表达，并且恢复了GPX4（P=0.029）表达；在HDM诱导的哮喘小鼠模型中，H&E染色和肺泡灌洗细胞计数显示，相较于HDM组，HDM+VBIT-4组的气道炎症细胞浸润减少，炎症细胞数量显著减少（P=0.0002）；瑞氏-姬萨姆染色也表明VBIT-4减少了肺泡灌洗液中嗜酸粒细胞的数量（P=0.001）免疫组织化学染色显示气道上皮细胞GPX4表达在VBIT-4干预下上调。结论 VDAC1参与HDM诱导的支气管哮喘的慢性气道炎症，可能通过引起铁死亡实现的。

关键词: 哮喘；铁死亡；线粒体；VDAC1

Abstract: Objective To investigate the role of voltage-dependent anion-selective channel protein 1 (VDAC1) in house dust mite (HDM)-induced asthmatic airway inflammation and its mechanism for regulating ferroptosis in airway epithelial cells. Methods Human airway epithelial (HBE) cells were exposed to a concentration gradient (200, 400 and 800 U) of HDM alone or in combination with treatment with 10 μmol/L VBIT-4 (a VDAC1 inhibitor) for 24 h, and the expressions of VDAC1 and ferroptosis-associated proteins in the cells were examined. Adult male BALB/c mice were treated with intranasal instillation of VBIT-4, HDM, or both, and the level of airway inflammation and the expressions of ferroptosis-associated proteins were detected with immunohistochemistry. Results In HBE cells, HDM exposure caused a significant increase of mitochondrial ROS (mtROS) production and obviously decreased the mitochondrial membrane potential. The exposed cells showed obviously increased protein expressions of VDAC1 (P=0.005) and FTH1 (P=0.030) but decreased protein expression of GPX4 (P=0.015) and FTH1 (P=0.037), while the treatment with VBIT-4 repressed the expression of GPX4 (P=0.001) and inhibited the expression of VDAC1. In BALB/c mice, treatment with VBIT-4 significantly improved HDM-induced airway inflammation by reducing the number of inflammatory cells (P=0.029) in the airway and the number of eosinophils in the alveolar lavage fluid. Immunohistochemical staining showed that GPX4 expression in the airway epithelial cells was significantly increased after treatment with VBIT-4. Conclusions VDAC1 participates in HDM-induced chronic airway inflammation in bronchial asthma by causing ferroptosis of the airway epithelial cells.

Key words: asthma; ferroptosis ; mitochondria; voltage-dependent anion-selective channel protein 1