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Jayter Paula, Carolina Modulo, Marco Andrey Frade, Peter Reinach, Eduardo Rocha; Rosmarinic Acid Suppresses Responses By Fibroblasts Contributing To Fibrosis Following Glaucoma Filtration Surgery. Invest. Ophthalmol. Vis. Sci. 2013;54(15):4796.
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© ARVO (1962-2015); The Authors (2016-present)
Tenon’s fibroblast proliferation and myofibroblast differentiation are contributors to the excessive subconjunctival scarring that occurs frequently after glaucoma filtration surgery. Rosmarinic acid (RA) is a natural polyphenol that has anti-fibrotic activities, found in many herbs. We reported that RA was able to control neovascularization, but not fibrosis, related to healing process in an experimental glaucoma surgery model (ARVO, 2012). Thus, the objective of this study was to determine if RA could suppress proliferation and differentiation of rabbits Tenon’s capsule fibroblasts (RTF).
RTF obtained from New Zealand rabbits were cultured in Dulbecco's modified Eagle's medium (DMEM). The dose dependent effects of RA (i.e. 0.3, 1.0 and 3.0 μM, for 12 h) on third passage cell proliferation were determined with the MTT assay in triplicate after 48 h. Immunofluorescence of alpha smooth muscle actin (alpha-SMA) assessed myofibroblast differentiation. Real-time reverse transcription polymerase chain reaction evaluated type I alpha I collagen (COL1A1) gene expression.
Only 1.0 μM (-10.5%; p=0.035) and 3.0 μM (-24.1%; p=0.014) RA suppressed proliferation after 48 h. On the other hand, at all RA concentrations, myofibroblast differentiation declined by 18.4% to 32.5%, compared to control. Moreover, COL1A1 mRNA expression also declined with RA treatments (p = 0.002).
RA suppressed RTF proliferation and transdifferentiation as well as COL1A1 gene expression. As RA treatment was previously associated with lower neovascularization in vivo, it remains an open question whether or not fibroblasts suppression by RA could control subconjunctival angiogenesis observed after glaucoma surgery.
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