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Peer-reviewed veterinary case report

Anti-fibrotic effects of lisinopril (ACE inhibitor) and fasudil (ROCK inhibitor) in combination for canine corneal fibrosis in vitro.

Journal:
Veterinary ophthalmology
Year:
2026
Authors:
Routh, Brayden L et al.
Affiliation:
Department of Veterinary Medicine and Surgery · United States
Species:
dog

Abstract

BACKGROUND: Corneal fibrosis is a leading cause of blindness in mammalian species and may result in compromised performance in sports and daily functions. This study evaluated the safety and anti-fibrotic effects of the FDA-approved drugs, angiotensin-converting enzyme inhibitor (ACE-I) lisinopril and rho-kinase inhibitor (ROCK-I) fasudil, alone and in combination, on the canine cornea using an established in&#xa0;vitro model. METHODS: To test the safety and efficacy of lisinopril and fasudil, primary canine corneal fibroblasts (CCFs) generated from donor corneas of healthy dogs (n&#x2009;=&#x2009;20) were used. A series of dose-dependent and time-dependent assays with lisinopril (1-50&#x2009;&#x3bc;M) and fasudil (1-10&#x2009;nM) were performed. qRT-PCR, immunofluorescence (IF) staining, cell viability assay, cell proliferation assay, LIVE/DEAD viability/cytotoxicity assay, TUNEL assay, and total cell count were performed. RESULTS: A 25-&#x3bc;M lisinopril and 3-nM fasudil dose were safe, nontoxic, and optimal for therapeutic evaluations in&#xa0;vitro. Treatments of lisinopril or fasudil, alone or in-combination, to CCFs grown in the presence of TGF-&#x3b2;1 (5&#x2009;ng/mL) showed inhibition of myofibroblast formation based on phase-contrast microscopy. The qRT-PCR and IF studies showed a significant decrease in expression of profibrotic markers, including &#x3b1;-smooth muscle actin (&#x3b1;-SMA; p&#x2009;<&#x2009;.0001), fibronectin (FN; p&#x2009;=&#x2009;.0002), tenascin C (TNC; p&#x2009;<&#x2009;.0001), Collagen I (Col-I; p&#x2009;<&#x2009;.0001), Collagen IIIA1 (Co-IIIA1; p&#x2009;<&#x2009;.0001), and Collagen IV (Co-lV; p&#x2009;<&#x2009;.0001). CONCLUSION: An ophthalmic formulation consisting of lisinopril and fasudil may offer a safe and effective method to treat canine corneal fibrosis. Additional studies evaluating safety and efficacy of this formulation in&#xa0;vivo are warranted.

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Original publication: https://pubmed.ncbi.nlm.nih.gov/39592228/