Yang Hua, Liu Mengzhu, Song Shiqi, Xu Qiang, Lee Jieun, Sun Jintao, Xue Shasha, Sun Xiaoyan, Che Chengye
Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao, China.
Department of Ophthalmology, School of Medicine, Pusan National University, Yangsan, Korea.
Invest Ophthalmol Vis Sci. 2025 Jun 2;66(6):32. doi: 10.1167/iovs.66.6.32.
This research was designed to explore the expression patterns and functional significance of hypoxia-inducible factor-1α (HIF-1α) in the inflammatory response associated with Aspergillus fumigatus (A. fumigatus) keratitis.
Mouse models of A. fumigatus keratitis were created by scraping the corneal epithelium and applying A. fumigatus on the corneal surface. In the in vitro experiments, human corneal epithelial cells (HCECs) and THP-1 macrophages stimulated by A. fumigatus were used to investigate the cellular responses. HIF-1α was inhibited using LW6. Western blot, immunofluorescence, and quantitative reverse transcription polymerase chain reaction (qRT-PCR) were performed to assess the expression levels of HIF-1α in A. fumigatus keratitis. The inflammatory response was evaluated using clinical scoring, corneal thickness measurements, hematoxylin and eosin (H&E) staining, corneal fluorescein sodium staining, and a cell scratch test. The polarization of macrophages was determined using flow cytometry. The molecular mechanisms of HIF-1α were assessed by qRT-PCR and Western blot.
In A. fumigatus keratitis, the expression of HIF-1α was significantly increased at both the mRNA and protein levels. Compared with the controls, HIF-1α inhibitor accelerated corneal epithelial repair, reduced the infiltration of macrophages, induced shift in macrophage polarization, and attenuated the inflammatory response. HIF-1α exerts a pro-inflammatory effect in A. fumigatus keratitis by modulating the expression of inflammatory mediators and engaging in pyroptosis via the caspase-8/GSDMD signaling pathway.
In conclusion, HIF-1α promotes A. fumigatus keratitis by inhibiting corneal epithelial repair and promoting inflammation, leading to increased severity of the disease.
本研究旨在探讨缺氧诱导因子-1α（HIF-1α）在烟曲霉性角膜炎相关炎症反应中的表达模式及功能意义。
通过刮除角膜上皮并在角膜表面接种烟曲霉建立烟曲霉性角膜炎小鼠模型。在体外实验中，使用经烟曲霉刺激的人角膜上皮细胞（HCECs）和THP-1巨噬细胞来研究细胞反应。使用LW6抑制HIF-1α。采用蛋白质免疫印迹法、免疫荧光法和定量逆转录聚合酶链反应（qRT-PCR）评估烟曲霉性角膜炎中HIF-1α的表达水平。通过临床评分、角膜厚度测量、苏木精-伊红（H&E）染色、角膜荧光素钠染色和细胞划痕试验评估炎症反应。使用流式细胞术确定巨噬细胞的极化。通过qRT-PCR和蛋白质免疫印迹法评估HIF-1α的分子机制。
在烟曲霉性角膜炎中，HIF-1α在mRNA和蛋白质水平上的表达均显著增加。与对照组相比，HIF-1α抑制剂加速了角膜上皮修复，减少了巨噬细胞浸润，诱导了巨噬细胞极化转变，并减轻了炎症反应。HIF-1α通过调节炎症介质的表达并经由半胱天冬酶-8/ Gasdermin D（caspase-8/GSDMD）信号通路参与细胞焦亡，在烟曲霉性角膜炎中发挥促炎作用。
总之，HIF-1α通过抑制角膜上皮修复和促进炎症反应来促进烟曲霉性角膜炎，导致疾病严重程度增加。
