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Neuroprotective effect of water-dispersible hesperetin in retinal ischemia reperfusion injury

  • Laboratory Investigation
  • Published:
Japanese Journal of Ophthalmology Aims and scope Submit manuscript

Abstract

Purpose

To determine whether water-dispersible hesperetin (WD-Hpt) can prevent degeneration of ganglion cell neurons in the ischemic retina.

Methods

Ischemia reperfusion (I/R) injury was induced by increasing the intraocular pressure of mice to 110 mmHg for 40 min. Mice received daily intraperitoneal injections with either normal saline (NS, 0.3 ml/day) or WD-Hpt (0.3 ml, 200 mg/kg/day). Reactive oxygen species (ROS) was assessed by dihydroethidium and nitrotyrosine formation. Inflammation was estimated by microglial morphology in the retina. Lipopolysaccharide (LPS)-stimulated BV-2 cells were used to explore the anti-inflammatory effect of WD-Hpt on activated microglia by quantifying the expression of IL-1β using real-time quantitative reverse transcription-polymerase chain reaction. Ganglion cell loss was assessed by immunohistochemistry of NeuN. Glial activation was quantified with glial fibrillary acidic protein (GFAP) immunoreactivity. Apoptosis was evaluated with a terminal deoxynucleotidyl transferase (TUNEL) assay and immunohistochemistry of cleaved caspase-3. Phosphorylation of extracellular signal-regulated kinase (p-ERK) was surveyed by western blotting.

Results

WD-Hpt decreased I/R-induced ROS formation. WD-Hpt alleviated microglial activation induced by I/R and reduced mRNA levels of IL-1β in LPS-stimulated BV-2. I/R resulted in a 37 % reduction in the number of ganglion cells in the NS-treated mice, whereas the reduction was only 5 % in the WD-Hpt-treated mice. In addition, WD-Hpt mitigated the immunoreactivity of GFAP, increased expression of cleaved caspase-3, increased number of TUNEL positive cells and p-ERK after I/R.

Conclusions

WD-Hpt protected ganglion cells from I/R injury by inhibiting oxidative stress and modulating cell death signaling. Moreover, WD-Hpt had an anti-inflammatory effect through the suppression of activated microglia.

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Acknowledgments

This work is supported by a Grant in aid for Young Scientists (B) Grant 25861609 (HY) from the Ministry of Education, Science and Culture, Tokyo, Japan.

Conflicts of interest

A. Shimouchi, Grant (Ezaki Glico Co., Ltd.); H. Yokota, Grant (Ezaki Glico Co., Ltd.); S. Ono, None; C. Matsumoto, None; T. Tamai, Employee (Ezaki Glico Co., Ltd.); H. Takumi, Employee (Ezaki Glico Co., Ltd.); S. P. Narayanan, None; S. Kimura, None; H. Kobayashi, None; R. B. Caldwell, None; T. Nagaoka, None; A. Yoshida, None.

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Authors and Affiliations

  1. Department of Ophthalmology, Asahikawa Medical University, Midorigaoka Higashi 2-1-1-1, Asahikawa, 078-8510, Japan

    Akito Shimouchi, Harumasa Yokota, Shinji Ono, Chiemi Matsumoto, Taiji Nagaoka & Akitoshi Yoshida

  2. Division of Immune Pathology, Department of Pathology, Asahikawa Medical University, Asahikawa, Japan

    Shoji Kimura & Hiroya Kobayashi

  3. Institute of Health Sciences, Ezaki Glico Co., Ltd, Osaka, Japan

    Toshihiro Tamai & Hiroko Takumi

  4. Vascular Biology Center, Georgia Regents University, Augusta, GA, USA

    Subbadra P. Narayanan & Ruth B. Caldwell

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  1. Akito Shimouchi
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  2. Harumasa Yokota
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  3. Shinji Ono
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Correspondence to Harumasa Yokota.

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Shimouchi, A., Yokota, H., Ono, S. et al. Neuroprotective effect of water-dispersible hesperetin in retinal ischemia reperfusion injury. Jpn J Ophthalmol 60, 51–61 (2016). https://doi.org/10.1007/s10384-015-0415-z

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  • DOI: https://doi.org/10.1007/s10384-015-0415-z

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