The neuroprotective effect of hesperidin in NMDA-induced retinal injury acts by suppressing oxidative stress and excessive calpain activation

Abstract

We found that hesperidin, a plant-derived bioflavonoid, may be a candidate agent for neuroprotective treatment in the retina, after screening 41 materials for anti-oxidative properties in a primary retinal cell culture under oxidative stress. We found that the intravitreal injection of hesperidin in mice prevented reductions in markers of the retinal ganglion cells (RGCs) and RGC death after N-methyl-D-aspartate (NMDA)-induced excitotoxicity. Hesperidin treatment also reduced calpain activation, reactive oxygen species generation and TNF-α gene expression. Finally, hesperidin treatment improved electrophysiological function, measured with visual evoked potential, and visual function, measured with optomotry. Thus, we found that hesperidin suppressed a number of cytotoxic factors associated with NMDA-induced cell death signaling, such as oxidative stress, over-activation of calpain, and inflammation, thereby protecting the RGCs in mice. Therefore, hesperidin may have potential as a therapeutic supplement for protecting the retina against the damage associated with excitotoxic injury, such as occurs in glaucoma and diabetic retinopathy.

Deleterious Effect of NMDA Plus Kainate on the Inner Retinal Cells and Ganglion Cell Projection of the Mouse

“…Since the doses administered to induce retinal excitotoxicity injury vary according to the literature used [17,[36][37][38][39][40][41], a series of experiments were performed to determine the ideal dose of joint administration of KA and NMDA in our model. Initially, in a series of mice (n = 3-4), different individual concentrations of NMDA (10, 30 and 100 mM) and KA (5, 10 and 20 mM) were administered by intraocular injection (see above), and the density of surviving RGC was estimated.…”

The Effect of Dietary Antioxidant Supplementation in Patients with Cataract and Glaucoma

“…Harris Here, we gave a daily dietary antioxidant supplement over 8 weeks to patients with cataract and glaucoma. The supplement was developed and rst reported by Maekawa et al, who identi ed the components of the supplement through screening, and found that it had a neuroprotective effect in a mouse glaucoma model [22]. We believe that the ndings reported here will shed light on the potential of new treatments for cataract and glaucoma based on antioxidant supplementation.…”

Ecel1 Knockdown With an AAV2-Mediated CRISPR/Cas9 System Promotes Optic Nerve Damage-Induced RGC Death in the Mouse Retina

“…A 2-lL NMDA solution diluted with PBS (15 mM) was injected into the vitreous cavity, as previously described. 24,25 In the controls that received NMDA injection, the same volume (2 lL) of PBS was injected. Vinblastine administration was also performed as described previously.…”

“…Transcriptional gene expression analysis was performed as previously described. 25 Briefly, retinal total RNA was isolated with a purification kit (miRNeasy Mini Kit; Qiagen, Inc., Hilden, Germany), and cDNA was synthesized with reverse transcriptase (SuperScript III; Life Technologies, Inc., Carlsbad, CA, USA). Quantitative PCR (qPCR) was performed on a realtime PCR system (7500 Fast Real-Time PCR System; Life Technologies, Inc.) using a universal master mix (TaqMan Fast Universal PCR Master Mix; Life Technologies, Inc.).…”

“…The electrophysiological response of the visual cortex was measured with the visual evoked potential (VEP), using a slight modification of a method we have previously described. 25 Under anesthesia induced with the intraperitoneal administration of a mixture of ketamine (37.5 mg/kg) and medetomidine (0.625 mg/kg), overnight dark-adapted mice were used to measure the photopic VEP with a Ganzfeld dome, as well as an acquisition system and an LED stimulator (PuREC and LS-100, respectively; all from Mayo Corp., Inazawa, Japan).…”