The evaluation of human tenon's fibroblasts and endothelial cell responses to antifibrotics alone and in combination with α-tocopherol

Curr Eye Res. 2015 Jan;40(1):19-29. doi: 10.3109/02713683.2014.909496. Epub 2014 Apr 21.

Abstract

Purpose: We aimed to evaluate the influence of current antifibrotic agents as well as the possible results obtained by combining these agents. This study included α-tocopherol, a strong antifibrotic and an efficient neuromediator of pathways used by other agents.

Materials and methods: Mitochondrial Bcl-2, Bax, cytochrome c and cytoplasmic caspase-3 expression, as well as toxic effect patterns, mitosis and cellular reactions due to α-tocopherol alone or combined with paclitaxel, mitomycin C and 5-flurouracil (5-FU), was studied in series obtained from human endothelial and primary Tenon's fibroblast cell cultures.

Results: The strongest apoptotic effect in both cell groups belonged to paclitaxel, followed by mitomycin C, and despite the overall suppressive effect of the α-tocopherol combination, mitomycin C increased its efficiency on the endothelial cells. The apoptosis/necrosis ratio was highest in α-tocopherol and lowest in paclitaxel, with α-tocopherol generally decreasing necrosis. Bax was observed at a high level with mitomycin C. Cytotoxicity was the highest with paclitaxel, and the caspase-3 reaction was markedly higher with mitomycin C in both cell types. In the α-tocopherol and 5-FU slides, mitosis and a layered formation were observed. The addition of α-tocopherol reduced the cytotoxicity of all antifibrotic agents in both cell series by decreasing the cell numbers, leading to necrosis.

Conclusions: Alone or in combination, the use of α-tocopherol and 5-FU is safer than other agents. By suppressing the cytotoxic effects of other antifibrotic agents, α-tocopherol is a promising drug for improving the effects of antifibrotics in many aspects of medicine. In addition, it has the potential to play a role beyond its antioxidant and antifibrotic activity in ocular surgery.

Keywords: 5-fluorouracil; Tenon's fibroblast; antifibrotic; apoptosis; endothelium; glaucoma surgery; mitomycin; α-tocopherol.

MeSH terms

  • Alkylating Agents / pharmacology*
  • Antioxidants / pharmacology*
  • Apoptosis / drug effects
  • Caspase 3 / metabolism
  • Cells, Cultured
  • Cytochromes c / metabolism
  • Drug Combinations
  • Fibroblasts / drug effects*
  • Fibroblasts / metabolism
  • Fibroblasts / pathology
  • Flow Cytometry
  • Fluorouracil / pharmacology
  • Human Umbilical Vein Endothelial Cells / drug effects*
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Human Umbilical Vein Endothelial Cells / pathology
  • Humans
  • Mitomycin / pharmacology
  • Necrosis
  • Paclitaxel / pharmacology
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Tenon Capsule / cytology*
  • alpha-Tocopherol / pharmacology*
  • bcl-2-Associated X Protein / metabolism

Substances

  • Alkylating Agents
  • Antioxidants
  • BAX protein, human
  • Drug Combinations
  • Proto-Oncogene Proteins c-bcl-2
  • bcl-2-Associated X Protein
  • Mitomycin
  • Cytochromes c
  • Caspase 3
  • alpha-Tocopherol
  • Paclitaxel
  • Fluorouracil