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(Investigative Ophthalmology and Visual Science. 2004;45:4226-4232.)
© 2004 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.04-0450
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Heme Oxygenase-1 Induced in Müller Cells Plays a Protective Role in Retinal Ischemia–Reperfusion Injury in Rats

Satoko Arai-Gaun,1,2 Naomichi Katai,1,2 Takanobu Kikuchi,3 Toru Kurokawa,1 Kouichi Ohta,1 and Nagahisa Yoshimura1

1From the Department of Ophthalmology and the 3Research Support Center for Human and Environmental Sciences, Shinshu University School of Medicine, Matsumoto, Japan.

PURPOSE. To investigate the protective roles played by heme oxygenase (HO)-1 and -2 in the rat retina after ischemia–reperfusion injury.

METHODS. Retinal ischemia was induced in rats by increasing the intraocular pressure to 110 mm Hg for 60 minutes. The expression of HO-1 and -2 in the retina was determined by Western blot, real-time polymerase chain reaction (PCR), and immunohistochemistry. To inhibit the upregulation of HO-1, short interfering (si)RNA of HO-1 was injected intravitreally before ischemia and that of green fluorescent protein (GFP) was used as the control. Müller cell damage was assessed by counting the number of S-100–positive cells. The number of macrophages invading the retina was determined by counting the number of ED-1–positive cells.

RESULTS. The expression of HO-1 mRNA and protein was upregulated at 6 hours after reperfusion and peaked at 12 to 24 hours, whereas that of HO-2 was not altered. HO-1 immunoreactivities were detected in Müller cells at 24 hours after reperfusion, and HO-2 immunoreactivities were detected in retinal cells. The HO-1 expression in the retina treated with siRNA of HO-1 was reduced at 12 and 24 hours after reperfusion compared with that injected with siRNA of GFP. The number of S-100–positive cells at 24 hours after reperfusion decreased significantly in retinas treated with HO-1 siRNA (P < 0.01). The number of macrophages that had infiltrated the retina was increased in retinas pretreated with the siRNA of HO-1 compared with those treated with siRNA of GFP. On day 14 after reperfusion, HO-1 siRNA-treated retinas showed severe retinal injury and destruction of the retinal architecture.

CONCLUSIONS. HO-1 promotes the survival of Müller cells after ischemia–reperfusion injury. Because inhibition of the upregulation of HO-1 resulted in an infiltration of inflammatory cells and destruction of the retina, the authors conclude that HO-1 induced in Müller cells plays a protective role in retinal ischemia-reperfusion.




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