Protective effect of senegenin on retinal ganglion cells in oxidative stress induced injury |
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Authors: | BIE Man XU Ying HU Hui-ling LU Dan ZHU Li-hong QI Ren-bin WANG Hua-dong LU Da-xiang |
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Affiliation: | 1. Department of Pathophysiology, Institute of Brain Research, Key Laboratory of State Administration of Traditional Chinese Medicine, School of Medicine, Jinan University,;2. Joint Laboratory for Brain Function and Health, Jinan University and The University of Hong Kong, Guangzhou 510632, China;3. Zhongshan Ophthalmological Center, Sun Yat-sen University, Guangzhou 510060, China |
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Abstract: | AIM: To evaluate the effect of senegenin (Sen) on H2O2-treated retinal ganglion cells (RGCs) and to explore its underlying mechanisms. METHODS: RGCs were retrograde labeled by injection of fluorogold into the superior colliculi of SD rats on the postnatal day 3. On the postnatal days 6 to 8, the retinas were dissociated with papain and cultured. Primary RGCs cultured in vitro were treated with H2O2 and/or various doses of Sen. The viability of RGCs was evaluated by counting the fluorescence-labeled neurons under microscope. The morphological changes of the nuclei in the retinal neurons were observed by Hoechst 33258 staining. Western blotting was applied to determine the expression of cleaved caspase-3, cytochrome C and Bcl-2 in cultured retinal neurons. RESULTS: Compared with the control cells, Sen at doses of 10, 20 or 40 μmol/L had no toxicity to RGCs (P>0.05). However, Sen at doses of 80 and 160 μmol/L had significant toxicity to RGCs (P<0.01). Compared with H2O2-injured group, Sen at doses of 10, 20 and 40 μmol/L effectively protected against H2O2-induced injury in RGCs (P<0.05) with the best efficiency at 40 μmol/L. Hoechst 33258 staining showed that the neuronal apoptosis caused by H2O2 was reduced by Sen. The results of Western blotting showed an up-regulation of Bcl-2, and decreased cytochrome C and cleaved caspase-3 levels by Sen in H2O2-treated retinal neurons. CONCLUSION: Sen is able to protect RGCs from H2O2-induced injury by enhancing Bcl-2 expression and inhibiting cell apoptosis. |
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Keywords: | Retinal ganglion cells Senegenin Oxidative stress |
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