Retinal Toxicity of Commercial Tissue Plasminogen Activator Is Mediated by the Induction of Nitric Oxide in the Mouse Retinal Primary Cells

Abstract

Purpose: Tissue plasminogen activator (tPA) is an efficient thrombolytic agent, but the dose-dependent retinal toxicity of intravitreal injection of commercial tPA (containing l-arginine) has been reported. Here, we sought to investigate the mechanism of tPA-induced cell death in mouse retinal cell cultures and the role of nitric oxide (NO). Methods: Primary retinal cell cultures were maintained using glial conditioned medium (GCM) solution. Mouse retinal cell death was observed by using Hoechst-propidium iodide staining. Mouse retinal cell death was also measured by lactate dehydrogenase (LDH) assay. The formation of NO was measured using Griess reagent. Results: tPA-induced cell death was detected in mouse retinal cell cultures by Hoechst-propidium iodide staining or LDH assay. l-Arginine seems to be the major factor in retinal toxicity of commercial tPA (containing l-arginine). The formation of NO was markedly increased in mouse retinal cell cultures treated with tPA (containing l-arginine) or l-arginine. NO inhibitor reduced the cell death induced by commercially available tPA or l-arginine. Conclusions: This study suggests that l-arginine from commercial tPA (containing l-arginine) induces the majority of cell death in mouse retinal cell cultures and that its cytotoxicity may depend on the induction of NO.

KEYWORDS:

• l-arginine
• nitric oxide
• retinal cell death
• tissue plasminogen activator
• toxicity