Effect of Amiloride to Retinal Toxicity Induced by Tissue Plasminogen Activator

PURPOSE: The effects of amiloride on cellular toxicity caused by tissue plasminogen activator (tPA) in mouse primary retinal cells were investigated. METHODS: Primary retinal cell cultures were maintained using glial conditioned medium. Commercial tPA and L-arginine were added, and the level of cyclic guanosine monophosphate (cyclic-GMP) in the culture supernatant was assessed using an ELISA assay. We measured the cell viability of cultured retinal cells pretreated with three different concentrations of amiloride (1, 10, and 100 microm) in addition to commercial tPA or L-arginine treatment. RESULTS: After exposing the cultured mouse retinal cells to tPA plus L-arginine or L-arginine alone, cyclic-GMP concentrations were 61.9 +/- 5.1 pmole/mL and 63.1 +/- 6.1 pmole/mL, respectively. However, the control group had a significantly lower concentration of cyclic-GMP (37.2 +/- 3.4 pmole/mL, p < 0.01). The cyclic GMP-dissolved solution did not cause retinal cell death. In the control group and the group treated with 1 microm amiloride and tPA containing L-arginine, the cell viability was 43.7% and 44.5%, respectively. However, cell viability increased to 70.6% with 10 microm amiloride and 78.4% with 100 microm amiloride (p = 0.015). CONCLUSIONS: L-arginine increases intracellular cyclic-GMP and may give rise to retinal cells through this mechanism. In addition, amiloride in concentrations greater than 10 microm protects against L-arginine-induced retinal cell death.

Resolution of experimental intravitreal fibrin by tissue plasminogen activator

Intravitreal fibrin clots were produced by intravitreal injection of 0.2 ml of autologous plasma in 62 rabbit eyes. The intravitreal injection of 0.25 micrograms or more of tissue plasminogen activator(tPA) resulted in a total clearing of intravitreal fibrin within one day in all treated eyes. This was significantly faster than in the control eyes, in which complete clearing was not seen until 8 days later. This represents the plateau on the dose-response curve in doses ranging from 0.25 to 200 micrograms. With light microscopy and transmission electron microscopy, retinal toxicity was demonstrated in eyes enucleated seven days after injection of 25 micrograms or more of tPA. This study demonstrates that tPA was effective and safe at 12.5 micrograms or less in clearing intravitreal fibrin in an experimental model. These results suggest that low dosages of tPA, probably of 3 micrograms or less, may be useful in the treatment of severe postvitrectomy fibrin formation seen clinically.