Wang Ning, Li Yizhou, Wang Xiaolei, Yang Lingling, Zhang Jing, Cheng Jun, Jiang Xiaoyue, Qi Xia, Wei Chao, Zhou Qingjun, Li Ya, Li Suxia
Eye Institute of Shandong First Medical University, Qingdao Eye Hospital of Shandong First Medical University, Qingdao, China.
State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China.
Invest Ophthalmol Vis Sci. 2025 Jun 2;66(6):28. doi: 10.1167/iovs.66.6.28.
This study aimed to elucidate the role and mechanism of corneal nerves in regulating epithelial cell response against ferroptosis.
Denervated mouse models were established via surgical axotomy and capsaicin treatment. Monochlorobimane staining was employed to detect cellular glutathione (GSH) levels in the corneal epithelium, and real-time quantitative PCR and immunofluorescence staining were used to evaluate GSH-related gene expression in denervated models and corneas of patients with neurotrophic keratitis. Scanning electron microscopy was utilized to observe mitochondrial morphology in corneal epithelial cells. Ferroptosis inhibitor ferrostatin-1 was administered post-corneal scrape in capsaicin-treated mice, followed by transcriptomic sequencing. The p53 agonist Kevetrin activated p53 in scraped corneas and cultured corneal epithelial cells. Furthermore, capsaicin was topically applied to Trp53+/- mice, followed by corneal epithelial scraping.
In denervated models, the expression of GSH-related genes was downregulated, and mitochondrial morphology exhibited characteristics of ferroptosis in corneal epithelial cells. The delay in corneal wound healing induced by TRPV1+ sensory denervation was ameliorated by ferrostatin-1 treatment. RNA sequencing and immunofluorescence staining demonstrated upregulated p53 in TRPV1-denervated mice, which was subsequently downregulated following ferrostatin-1 treatment. Kevetrin exacerbated wound healing delays, whereas Trp53+/- mice exhibited accelerated healing post-capsaicin denervation compared to wild-type controls.
TRPV1+ sensory nerves play a regulatory role in preventing ferroptosis of corneal epithelial cells through the p53/AKT/mTOR signaling pathway. Targeting this pathway may offer therapeutic potential for neurotrophic keratopathy and related disorders.
本研究旨在阐明角膜神经在调节上皮细胞抗铁死亡反应中的作用及机制。
通过手术切断轴突和辣椒素处理建立去神经支配小鼠模型。采用单氯双氢杨梅素染色检测角膜上皮细胞内的谷胱甘肽（GSH）水平，运用实时定量聚合酶链反应和免疫荧光染色评估去神经支配模型及神经营养性角膜炎患者角膜中GSH相关基因的表达。利用扫描电子显微镜观察角膜上皮细胞中的线粒体形态。在辣椒素处理的小鼠角膜刮伤后给予铁死亡抑制剂铁抑素-1，随后进行转录组测序。p53激动剂凯维菌素在刮伤的角膜和培养的角膜上皮细胞中激活p53。此外，将辣椒素局部应用于Trp53+/-小鼠，随后进行角膜上皮刮伤。
在去神经支配模型中，GSH相关基因的表达下调，角膜上皮细胞中的线粒体形态呈现铁死亡特征。铁抑素-1治疗改善了由TRPV1+感觉神经去神经支配引起的角膜伤口愈合延迟。RNA测序和免疫荧光染色显示TRPV1去神经支配小鼠中p53上调，而在铁抑素-1治疗后p53随后下调。凯维菌素加剧了伤口愈合延迟，而与野生型对照相比，Trp53+/-小鼠在辣椒素去神经支配后表现出加速愈合。
TRPV1+感觉神经通过p53/AKT/mTOR信号通路在预防角膜上皮细胞铁死亡中发挥调节作用。靶向该通路可能为神经营养性角膜病及相关疾病提供治疗潜力。
