Conclusions
VDAC1 participates in HDM-induced chronic airway inflammation in bronchial asthma by causing ferroptosis of the airway epithelial cells. 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。
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.
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.
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. 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。