Verteporfin combined with ROCK inhibitor promotes the restoration of corneal endothelial cell dysfunction in rats

• Published in: Biochemical pharmacology 2025-01, Vol.231, p.116641, Article 116641
• Main Authors: Zhang, Xue, Liu, Hongling, Wan, Chao, Li, Yijian, Ren, Chunge, Lu, Jia, Liu, Yong, Yang, Yuli
• Format: Article
• Language: English
• Subjects: Amides - pharmacology; Animals; Aquaporin 1 - antagonists & inhibitors; Aquaporin 1 - genetics; Aquaporin 1 - metabolism; Corneal endothelial cell dysfunction; Corneal endothelial-to-mesenchymal transition; Drug Therapy, Combination; Endothelial Cells - drug effects; Endothelial Cells - metabolism; Endothelium, Corneal - drug effects; Endothelium, Corneal - metabolism; Endothelium, Corneal - pathology; Humans; Male; Photosensitizing Agents - pharmacology; Photosensitizing Agents - therapeutic use; Protein Kinase Inhibitors - pharmacology; Pyridines - pharmacology; Rats; Rats, Sprague-Dawley; rho-Associated Kinases - antagonists & inhibitors; rho-Associated Kinases - metabolism; Verteporfin; Verteporfin - pharmacology; Verteporfin - therapeutic use; YAP-Signaling Proteins - metabolism; Yes-associated protein
• ISSN: 0006-2952; 1873-2968; 1873-2968
• DOI: 10.1016/j.bcp.2024.116641

[Display omitted] Corneal endothelial cells (CECs) dysfunction frequently results in a hazy, edematous cornea due to corneal endothelial decompensation and is a major cause of corneal blindness. Drug interventions provide a less invasive alternative to corneal transplantation surgery. However, endothelial-to-mesenchymal transition (EndMT) limits CECs function. Rho-kinase (ROCK) inhibitors, shown in numerous studies to be an adjunctive therapy for CECs dysfunction, cannot completely reverse pathological EndMT caused by inflammatory environmental damage. Verteporfin (VP) is an inhibitor of Yes-associated protein (YAP) and has significant inhibitory effects on cell fibrosis and mesenchymal transition. Here, we explored VP’s utility in mitigating EndMT during ROCK inhibitors treatment of corneal endothelial dysfunction. We surgically constructed a rat model of CECs injury and studied VP and ROCK inhibitors’ effects on EndMT, cell proliferation, and corneal edema using RNA-Seq sequencing, immunofluorescence, optical coherence tomography, and qPCR. The results indicated that YAP expression in human fetal CECs was higher than in adults and decreased with age in rats. Moreover, YAP expression in human CECs was negatively correlated with functional genes, such as AQP1 and ATP1A1. VP effectively reversed EndMT and accelerated corneal hydration regression. However, it inhibited CECs proliferation. We also confirmed that the optimal ratio of VP combined with Y-27632 (ROCK inhibitor) was 1:1, promoting CECs proliferation and reversing EndMT by down-regulating transcription factors downstream of TGF-β signaling, thereby increasing CECs functional and intercellular adhesion proteins. These combined effects promote corneal endothelial damage repair, providing a new treatment strategy.