Abstract
Purpose: :
to study the possible mechanisms of action involved in the anti-angiogenic effects observed with NOV C-ter (Sisène, France) in the corneal micropocket assay.
Methods: :
The corneal micropocket assay was performed on 37 Lewis male rats of 6-8 weeks old. LPS pellets (Sigma-Aldrich, France) were implanted into the corneal stroma followed by sub-conjunctival injections every other day for 8 days. Rats were randomly assigned to the following groups: no treatment, PBS, bevacizumab 250µg, NOV C-ter 5 µg, NOV C-ter 10 µg and NOV 2µg. At day 9, angiogenesis was assessed clinically using a grading score, animals were euthanized and corneas and conjunctivas were extracted. Real-time PCR analysis was performed to evaluate the expression of several genes: VEGF, VEGF-R1, VEGF-R2, CD31, iNOS, IL-1β, IL-6, TNF-α, MCP-1, desmin, STAT3 and connexin-43.
Results: :
NOV C-ter 10µg and NOV 2µg significantly reduced LPS induced angiogenesis (P < 0.05). No significant effect was observed for bevacizumab. Real-time PCR analysis of the corneas showed no significant effects on the expression of VEGF, VEGF-R1, VEGF-R2, iNOS, IL-1β, MCP-1, desmin, and STAT3. IL-6 was downregulated by NOV 2µg and NOV C-ter 10µg (P < 0.0001), CD31, TNF-α and connexin-43 were down-regulated by NOV 2µg (P < 0.05). In the conjunctivas, NOV C-ter 10µg down-regulated iNOS, MCP-1, connexin-43 and STAT3 (P < 0.05).
Conclusions: :
Anti-angiogenic effects observed for NOV C-ter, in the rat corneal micropocket assay, seem to be independent of the VEGF pathway and may linked to down-regulation of inflammation mediators.
Keywords: neovascularization • drug toxicity/drug effects • inflammation