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From the Department of Ophthalmology, Shinshu University School of Medicine, Matsumoto, Japan.
PURPOSE. To evaluate possible roles of caspase-1 and caspase-3 in retinal ischemia–reperfusion injury.
METHODS. Retinal ischemia was induced in rats by increasing the intraocular pressure to 110 mm Hg for 60 minutes. Expression of caspase-1 and caspase-3 was studied at the mRNA and protein levels using immunohistochemical staining, western blot analysis, semiquantitative reverse transcription–polymerase chain reaction, and assay of the enzymatic activities. Apoptotic retinal neurons were detected by the TdT-dUTP terminal nick-end labeling (TUNEL) method. To study the roles of the caspases in retinal ischemia–reperfusion injury, an inhibitor of caspase-1, acetyl-tyrosyl-valyl-alanyl-aspart-1-al (Ac-YVAD-CHO; total dose, 10-7 moles) and that of caspase-3, acetyl-aspartyl-glutamyl-valyl-aspart1-al (Ac-DEVD-CHO; total dose, 10-7 moles) was injected intravitreally and the number of TUNEL-positive cells was compared with the number in sections not treated with the inhibitors.
RESULTS. In the inner nuclear layer (INL), caspase-3-like immunoreactivity was predominantly detected, whereas caspase-1-like immunoreactivity was more predominant in the outer nuclear layer (ONL). Expression of caspase-1 and -3 was upregulated at the protein and gene levels 24 hours after reperfusion. Intravitreal injection of Ac-DEVD-CHO decreased the number of TUNEL-positive cells more significantly in the INL than in the ONL (P < 0.01) at 24 hours, whereas, intravitreal injection of Ac-YVAD-CHO was more effective in decreasing the number in the ONL (P < 0.05).
CONCLUSIONS. These findings suggest a possibility that cell-type–specific activation of caspases takes place in retinal ischemia–reperfusion injury, and such caspase may induce retinal neuronal cell death.
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