Suberoylanilide hydroxamic acid (vorinostat): its role on equine corneal fibrosis and matrix metalloproteinase activity.
Vet Ophthalmol
; 17 Suppl 1: 61-8, 2014 Jul.
Article
in En
| MEDLINE
| ID: mdl-25126665
ABSTRACT
OBJECTIVE:
To explore the effect of suberoylanilide hydroxamic acid (SAHA) (i) on corneal fibroblast differentiation, morphology, and viability; and (ii) on the expression levels of matrix metalloproteinases (MMPs) 2 and 9 using an in vitro model of equine corneal fibrosis. PROCEDURE Healthy donor corneas were used to generate primary cultures of equine corneal fibroblasts. The fibroblasts were exposed to 5 ng/mL TGFß1 to induce myofibroblast formation. The cultures were treated with either 5 µm or 10 µm SAHA for 72 h in the presence of TGFß1. Real-time PCR and immunocytochemistry were used to determine the antifibrotic efficacy of SAHA by quantifying α-smooth muscle actin (αSMA), a marker of myofibroblast formation and fibrosis. Real-time PCR was used to determine the effects of SAHA on MMP2 and MMP9 expression. Cytotoxicity of SAHA was evaluated with phase contrast microscopy and trypan blue exclusion assays.RESULTS:
Suberoylanilide hydroxamic acid (SAHA) significantly attenuated TGFß1-induced differentiation of equine fibroblasts to myofibroblasts as indicated by 3- to 3.5-fold (P < 0.001) decrease in αSMA mRNA and 86-88% (P < 0.001) decrease in αSMA+ immunocytochemical staining. SAHA treatment also resulted in 4.5- to 5.5-fold (P < 0.01) decrease in MMP9 expression. A dose-dependent bimodal effect of SAHA on MMP2 expression was noted (3.5-fold increase with 5 µm dose; 0.5-fold decrease with 10 µm dose). No change in fibroblast viability was observed with a 5 µm SAHA dose, whereas a 10 µm dose resulted in a moderate 17% decrease in cell viability.CONCLUSIONS:
Suberoylanilide hydroxamic acid (SAHA) can effectively inhibit TGFß-induced differentiation of equine corneal fibroblasts to myofibroblasts and modulates MMP production in vitro.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Cicatrix
/
Matrix Metalloproteinases
/
Histone Deacetylase Inhibitors
/
Corneal Injuries
/
Horse Diseases
/
Hydroxamic Acids
Limits:
Animals
Language:
En
Journal:
Vet Ophthalmol
Journal subject:
MEDICINA VETERINARIA
/
OFTALMOLOGIA
Year:
2014
Document type:
Article