SIRT1 is required for the neuroprotection of resveratrol on retinal ganglion cells after retinal ischemia-reperfusion injury in mice
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Retinal ganglion cells (RGCs) loss is closely related to visual impairment in glaucoma, so the neuroprotection on RGCs is important and novel for glaucoma research. SIRT1, a family member of sirtuins, is implicated in many crucial processes of eye diseases. The purpose of this study is to determine the neuroprotection of SIRT1 on RGCs and to investigate the underlying mechanisms of these effects in an experimental model for acute glaucoma.
Retinal ischemia-reperfusion (IR) injury was induced in C57BL/6J mice. Resveratrol (RSV, activator of SIRT1) and sirtinol (inhibitor of SIRT1) were injected intravitreally 1 day before IR injury. RGCs survival rate was quantified by immunofluorescence staining. RGCs apoptosis was evaluated by the staining of TUNEL and cleaved caspase-3, and SIRT1 level was detected by western blot. Expressions of phospho-Akt, Akt, Bax, and Bcl-2 were further determined by western blot to investigate the neuroprotective mechanisms of SIRT1.
RGCs survival rates and SIRT1 levels were decreased over time after IR injury. Intravitreal injection of RSV remarkably attenuated RGCs loss in a dose-dependent manner, and the most effective concentration of RSV was 100 μM. Up-regulation of SIRT1 by RSV significantly inhibited RGCs apoptosis, increased p-Akt level, decreased Bax and cleaved caspase-3 expressions, and all these effects were diminished by 100 μM sirtinol. Moreover, there were no significant changes in total Akt and Bcl-2 levels.
SIRT1 activation by RSV confers neuroprotection on RGCs in retinal IR injury through the activation of Akt pathway and subsequent suppression of mitochondrial apoptotic pathway. Determination of the effective concentration of intravitreal injection of RSV also provides a theoretical basis for the clinical application of RSV.
KeywordsSIRT1 Resveratrol Retinal ganglion cell Ischemia-reperfusion Apoptosis Neuroprotection
We thank Ying Li, Xinlan Lei, Qinqin Deng, and Xiao Zhang for their constructive scientific suggestions.
This work was funded by the National Natural Science Foundation of China (grant number 81271025).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
All applicable international, national, and institutional guidelines for the care and use of animals were followed. The article does not contain any studies with human participants performed by any of the authors.
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