Ghrelin, a 28-amino acid peptide hormone, has protective effects on neuronal cells. The present study aimed to examine the neuroprotective effects of ghrelin on the rat retinal ganglion cells in the rotenone-induced in vitro model of Parkinson's disease (PD). Cell viability and cell apoptosis were determined by MTT assay and flow cytometry, respectively. Mitochondrial functions were detected by mitochondrial complex I activity assay and mitochondrial membrane potential (MMP) assay. The mRNA and protein expression levels were determined by qRT-PCR and western blot, respectively. Rotenone significantly suppressed cell viability and increased cell apoptosis, also decreased the mitochondrial complex I activity as well as MMP in rat retinal ganglion cell line (RGC-5). Growth hormone secretagogue receptor (Ghsr) siRNA transfection significantly suppressed the expression of Ghsr in RGC-5 cells. Ghrelin treatment attenuated the effects of rotenone-induced changes in cell viability, cell apoptosis and mitochondrial functions in RGC-5 cells. Post-transcriptional suppression by Ghsr siRNA transfection and treatment with GHS-R antagonist, YIL781, both significantly attenuated the effects of ghrelin in RGC-5 cells. Rotenone decreased the protein levels of Bcl-2 and increased the protein levels of Bax, cleaved caspase-3 and cleaved caspase-9, and this effect was reversed by ghrelin treatment. Ghrelin also prevented the inhibitory effects of rotenone on the AKT-mTOR signaling. The effects of ghrelin on the rotenone-induced changes in apoptosis-related protein levels and AKT-mTOR signaling were attenuated by Ghsr siRNA transfection and treatment with YIL781 in the RGC-5 cells. In addition, both rapamycin and AKT inhibitor IV pre-treatment significantly attenuated the effects of ghrelin on rotenone-induced changes in cell viability and cell apoptosis. In conclusion, ghrelin by acting on the GSH-R to protect rat retinal ganglion cells against rotenone via inhibiting apoptosis and restore mitochondrial functions in RGC-5 cells, and this effect was partially associated with the AKT-mTOR signaling pathway in RGC-5 cells.
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