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Effects of docosahexaenoic acid on the viability of human tenon's fibroblasts
Author(s) -
De Lazzer A.,
Acar N.,
Bretillon L.,
Bron A.M.,
Creuzot Garcher C.
Publication year - 2017
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2017.04121
Subject(s) - docosahexaenoic acid , viability assay , fibroblast , mtt assay , wound healing , cytotoxicity , chemistry , mapk/erk pathway , p38 mitogen activated protein kinases , toxicity , fibrosis , blot , polyunsaturated fatty acid , pharmacology , cell growth , microbiology and biotechnology , cell , medicine , biochemistry , biology , fatty acid , pathology , surgery , kinase , in vitro , gene
Purpose Excessive scarring of the conjunctiva is the major cause of failure of the surgical treatment for glaucoma. The activation of human Tenon's fibroblast ( HTF s) is supposed to be responsible for wound healing and scar formation. Antimetabolites are used to inhibit subconjunctival fibrosis but these molecules may lead to serious complication. Previous studies have shown that omega‐3 polyunsaturated fatty acids ( PUFA s) such as docosahexaenoic acid ( DHA ) have anti‐inflammatory, anti‐proliferative and anti‐angiogenic properties. The aims of this study were to evaluate DHA as a potential inhibitor of human Tenon's fibroblasts. Methods Primary HTF cells were obtained from patients during glaucoma surgery and were cultured in DMEM medium. After stimulation with TGF ‐ β 2, HTF s were treated with MMC (0.01 mg/ mL ) and different concentrations of DHA (50, 100 and 200 μ M) for 48 hours. Lipid profile of HTF s was determined by gas chromatography, cell proliferation by Ki67 assay, cell migration was evaluated by videomicroscopy and cell toxicity by MTT assay. The expression α ‐ SM actin and MAPK p38, ERK , JNK in their non‐phosphorylated and phosphorylated forms was determined by western blotting. Results MMC reduced proliferation, migration compared to control but showed important toxicity suggested by the increased respiratory rate of mitochondrie in cytotoxicity assay and confirmed by a G2 arrest in cell cycle. DHA had no significant impact on cellular viability. DHA at 200 μ M showed an efficacity similar to MMC . A reduction of the ability of cells to migrate was observed with DHA in a dose‐dependent manner. DHA efficiently dicreased α ‐ SM actin expression of myofibroblasts. Proteins from the Smad signaling pathway could not be observed. Conclusions These findings indicate that DHA inhibits migration and differentiation of HTF s to myofibroblats.