PDF(Pubmed)
Abstract：
BACKGROUND: Corneal injuries, often leading to severe vision loss or blindness, have traditionally been treated with the belief that limbal stem cells (LSCs) are essential for repair and homeostasis, while central corneal epithelial cells (CCECs) were thought incapable of such repair. However, our research reveals that CCECs can fully heal and maintain the homeostasis of injured corneas in rats, even without LSCs. We discovered that CXCL14, under PAX6\'s influence, significantly boosts the stemness, proliferation, and migration of CCECs, facilitating corneal wound healing and homeostasis. This finding introduces CXCL14 as a promising new drug target for corneal injury treatment.
METHODS: To investigate the PAX6/CXCL14 regulatory axis\'s role in CCECs wound healing, we cultured human corneal epithelial cell lines with either increased or decreased expression of PAX6 and CXCL14 using adenovirus transfection in vitro. Techniques such as coimmunoprecipitation, chromatin immunoprecipitation, immunofluorescence staining, western blot, real-time PCR, cell colony formation, and cell cycle analysis were employed to validate the axis\'s function. In vivo, a rat corneal epithelial injury model was developed to further confirm the PAX6/CXCL14 axis\'s mechanism in repairing corneal damage and maintaining corneal homeostasis, as well as to assess the potential of CXCL14 protein as a therapeutic agent for corneal injuries.
RESULTS: Our study reveals that CCECs naturally express high levels of CXCL14, which is significantly upregulated by PAX6 following corneal damage. We identified SDC1 as CXCL14\'s receptor, whose engagement activates the NF-κB pathway to stimulate corneal repair by enhancing the stemness, proliferative, and migratory capacities of CCECs. Moreover, our research underscores CXCL14\'s therapeutic promise for corneal injuries, showing that recombinant CXCL14 effectively accelerates corneal healing in rat models.
CONCLUSIONS: CCECs play a critical and independent role in the repair of corneal injuries and the maintenance of corneal homeostasis, distinct from that of LSCs. The PAX6/CXCL14 regulatory axis is pivotal in this process. Additionally, our research demonstrates that the important function of CXCL14 in corneal repair endows it with the potential to be developed into a novel therapeutic agent for treating corneal injuries.
METHODS: To investigate the PAX6/CXCL14 regulatory axis\'s role in CCECs wound healing, we cultured human corneal epithelial cell lines with either increased or decreased expression of PAX6 and CXCL14 using adenovirus transfection in vitro. Techniques such as coimmunoprecipitation, chromatin immunoprecipitation, immunofluorescence staining, western blot, real-time PCR, cell colony formation, and cell cycle analysis were employed to validate the axis\'s function. In vivo, a rat corneal epithelial injury model was developed to further confirm the PAX6/CXCL14 axis\'s mechanism in repairing corneal damage and maintaining corneal homeostasis, as well as to assess the potential of CXCL14 protein as a therapeutic agent for corneal injuries.
RESULTS: Our study reveals that CCECs naturally express high levels of CXCL14, which is significantly upregulated by PAX6 following corneal damage. We identified SDC1 as CXCL14\'s receptor, whose engagement activates the NF-κB pathway to stimulate corneal repair by enhancing the stemness, proliferative, and migratory capacities of CCECs. Moreover, our research underscores CXCL14\'s therapeutic promise for corneal injuries, showing that recombinant CXCL14 effectively accelerates corneal healing in rat models.
CONCLUSIONS: CCECs play a critical and independent role in the repair of corneal injuries and the maintenance of corneal homeostasis, distinct from that of LSCs. The PAX6/CXCL14 regulatory axis is pivotal in this process. Additionally, our research demonstrates that the important function of CXCL14 in corneal repair endows it with the potential to be developed into a novel therapeutic agent for treating corneal injuries.
摘要：
背景：角膜损伤，通常导致严重的视力丧失或失明，传统上认为角膜缘干细胞（LSCs）对于修复和稳态至关重要，而中央角膜上皮细胞（CCEC）被认为无法进行这种修复。然而,我们的研究表明，CCEC可以完全治愈和维持大鼠受损角膜的稳态，即使没有LSC。我们发现CXCL14在PAX6的影响下，显著提高了干性，扩散,以及CCEC的迁移，促进角膜伤口愈合和体内平衡。这一发现将CXCL14作为角膜损伤治疗的一个有希望的新药靶点。
方法：为了研究PAX6/CXCL14调节轴在CCECs伤口愈合中的作用，我们使用腺病毒转染体外培养了PAX6和CXCL14表达增加或减少的人角膜上皮细胞系。技术，如免疫共沉淀，染色质免疫沉淀，免疫荧光染色，westernblot,实时PCR，细胞集落形成，和细胞周期分析用于验证轴的功能。在体内,建立了大鼠角膜上皮损伤模型，以进一步证实PAX6/CXCL14轴在修复角膜损伤和维持角膜稳态中的机制，以及评估CXCL14蛋白作为角膜损伤治疗剂的潜力。
结果：我们的研究表明，CCEC天然表达高水平的CXCL14，在角膜损伤后PAX6显著上调。我们鉴定SDC1为CXCL14受体，其接合激活NF-κB通路，通过增强干性刺激角膜修复，增殖性,和CCEC的迁移能力。此外，我们的研究强调了CXCL14对角膜损伤的治疗前景，表明重组CXCL14可有效加速大鼠模型的角膜愈合。
结论：CCECs在修复角膜损伤和维持角膜稳态中起着重要的独立作用。与LSC不同。PAX6/CXCL14调节轴在此过程中至关重要。此外,我们的研究表明,CXCL14在角膜修复中的重要功能使其有可能发展成为治疗角膜损伤的新型治疗剂.
方法：为了研究PAX6/CXCL14调节轴在CCECs伤口愈合中的作用，我们使用腺病毒转染体外培养了PAX6和CXCL14表达增加或减少的人角膜上皮细胞系。技术，如免疫共沉淀，染色质免疫沉淀，免疫荧光染色，westernblot,实时PCR，细胞集落形成，和细胞周期分析用于验证轴的功能。在体内,建立了大鼠角膜上皮损伤模型，以进一步证实PAX6/CXCL14轴在修复角膜损伤和维持角膜稳态中的机制，以及评估CXCL14蛋白作为角膜损伤治疗剂的潜力。
结果：我们的研究表明，CCEC天然表达高水平的CXCL14，在角膜损伤后PAX6显著上调。我们鉴定SDC1为CXCL14受体，其接合激活NF-κB通路，通过增强干性刺激角膜修复，增殖性,和CCEC的迁移能力。此外，我们的研究强调了CXCL14对角膜损伤的治疗前景，表明重组CXCL14可有效加速大鼠模型的角膜愈合。
结论：CCECs在修复角膜损伤和维持角膜稳态中起着重要的独立作用。与LSC不同。PAX6/CXCL14调节轴在此过程中至关重要。此外,我们的研究表明,CXCL14在角膜修复中的重要功能使其有可能发展成为治疗角膜损伤的新型治疗剂.
