Intravitreal Injection of Hydrogen Peroxide Induces Acute Retinal Degeneration, Apoptosis, and Oxidative Stress in Mice

Abstract EN

Purpose.

Oxidative stress is a common pathological condition for multiple retinal diseases.

Hydrogen peroxide (H2O2) has been applied as an oxidative stress inducer for the in vitro studies.

Here, we report the in vivo effect of H2O2 exposure to the mouse retina and its underlying mechanism.

Methods.

The H2O2 or saline solution was intravitreally injected into the eyes of female C57BL/6J mice for two consecutive days.

The retinal structure was evaluated by in vivo imaging using spectral domain optical coherence tomography (OCT) and validated by histological assessment as well as retinal marker expression.

In addition, retinal stress, cell apoptosis, and antioxidant enzyme expression were also determined.

Results.

Retinal and outer nuclear layer thickness thinning was observed at days 7 and 14 by OCT imaging with the treatment of 10 μg H2O2, which was confirmed by the histopathological analysis.

The expressions of photoreceptor (Rho, Rora, Rorb, and Rcvrn), bipolar cell (Chat and Calb2), and retinal pigment epithelial (Rpe65) markers were reduced in the H2O2-treated group, whereas the expression of retinal ganglion cell marker (Tubb3) was increased.

TUNEL-positive cells were obviously found in the outer nuclear layer and inner nuclear layer of H2O2-treated mice but sparely found in the ganglion cell layer.

Coherently, apoptotic gene expressions (Casp3, Casp9, Bax, and Parp8) were significantly increased in the retina with increasing dosages of H2O2, while Bcl2 expression was mildly decreased.

In addition, the expressions of Gfap and antioxidant enzyme genes (Txn2, Sod2, and Gpx4) were significantly upregulated in the retina after the H2O2 treatment, compared to the vehicle control group.

Conclusions.

This study revealed that intravitreal injection of H2O2 induces acute retinal damage by increasing oxidative stress and cell apoptosis in the retina.

This acute retinal degeneration mouse model could provide a platform for drug screening against oxidative stress and retinal diseases.