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¹Bağcılar E.A. Hastanesi, Göz Kliniği, İstanbul, Uzm. Dr.
²İ.Ü, İstanbul Tıp Fakültesi, Fizyoloji A.D., İstanbul, Prof. Dr.
³İ.Ü, Fen Fakültesi, Biyoloji Bölümü, İstanbul, Uzm. Dr.
⁴İ.Ü., İstanbul Tıp Fakültesi, Fizyoloji A.D., İstanbul, Asist. Dr.
⁵İ.Ü., Fen Fakültesi, Moleküler Biyoloji ve Genetik Bölümü, İstanbul, Uzm. Dr.
⁶İ.Ü., Fen Fakültesi, Biyoloji Bölümü, İstanbul, Asist. Dr.
⁷İ.Ü., Fen Fakültesi, Biyoloji Bölümü, İstanbul, Doç. Dr. Purpose: Side effects current antifibrotics on endothelium, limits their use in glaucoma surgery. The natural antioxidant, α- tocopherol is also a neuromediator, and it not only prevents tenon’s fibroblast proliferation and fibrocellular scar formation, but effects similar pathways with those of other antifibrotics as well. In this study, apoptotic effects of α-tocopherol and other antifibrotics -in concordant doses with their in vivo uses, on human endothelial cells were examined in vitro in various combinations and durations.
Materials and Methods: Effects of antifibrotic agents in ECV304 Human Umblical Vein Endothelial Cell Cultures were evaluated in single 5 minnutes applications and in combination with 10 days of α-tocopherol additions. Paclitaxel, mitomycin, 5-FU and α-tocopherol were used as antifibrotic agents. Apoptotic indexes were found with DAPI staining in fluorescence microscope. Apoptotic cells were demonstrated immunohistochemically in preperates stained with peroxidase labeled caspase 3 monoclonal antibodies and bcl-2 ve bax gene expressions were evaluated with PCR. Mitotic indexes were determined to evaluate the proliferative trends.
Results: Mitomycin and paclitaxel groups demonstrated a significant AI increase, which were augmented by α-tocopherol addition. Either bcl-2/bax ratio decrease, or caspase activity were found only in Mitomycin group and further augmentation was achieved with α-tocopherol addition. High proliferation was observed in tocopherol and paclitaxel+ tocopherol groups.
Conclusion: Combination with α-tocopherol did not reduce the apoptotic effects of antifibrotics on human endothelial cells. Single and combined uses of 5-FU and α-tocopherol have been found to be safe for their long and short term effects were not statistically different from controls. Keywords : Alpha-tocopherol, mitomycin, 5-FU, antifibrotic, apoptosis