Abstract
This study aimed to clarify the regulation role of miR-708 and miR-335-3p in retinal ganglion cell (RGC) autophagy and apoptosis in glaucoma. Chronic glaucoma mice were established by laser photocoagulation. RGCs were isolated and transfected with a series of plasmids and the cultured in 60 mmHg pressure. miR-335-3p, miR-708, and ATG3 mRNA expressions were detected by qRT-PCR. Protein levels of ATG3, autophagy-related protein LC3, and p62 were detected by Western blot. The apoptosis of RGCs was detected by flow cytometry. The regulation role of miR-335-3p/miR-708 in ATG3 was confirmed by the dual-luciferase reporter gene. The expressions of several miRNAs were measured in retinal tissues from chronic glaucoma mice and RGCs under pressure conditions, and results showed that both miR-335-3p and miR-708 were down-regulated. Besides, the inhibition of miR-708 and miR-335-3p induced the apoptosis of RGCs through promoting autophagy. Also, miR-708 and miR-335-3p could bind to ATG3 and targeted regulated ATG3. Furthermore, the interference with miR-708/miR-335-3p induced RGC apoptosis by up-regulating ATG3 to promote autophagy. In general, the down-regulation of miR-708 and miR-335-3p contributed to the apoptosis of RGCs through promoting autophagy in glaucoma.
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This work was funded by the National Natural Science Foundation of China (No. 81660734) and the Natural Science Foundation of Inner Mongolia (No. 2018MS08118).
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All animal experiments were approved by the Medical Ethics Committee of the Affiliated Hospital of Guangdong Medical University.
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Supplemental Figure 1
miR-708 and miR-335-3p regulated the expression of ATG3. mimic-NC, miR-708 mimic+miR-335-3p mimic, miR-708 mimic+miR-335-3p mimic+NC, miR-708 mimic+miR-335-3p mimic+WT-ATG3, miR-708 mimic+miR-335-3p mimic+MUT-ATG3 were transfected into RGCs, and then the cells were cultured in a 60 mmHg pressure incubator for 24 h. A. Detection of ATG3 mRNA level. B. Detection of ATG3 protein level. **p < 0.01 vs Pressure + mimic-NC or Pressure + miR-708 mimic+miR-335-3p mimic+NC. NC: Negative control. N = 3. (PNG 309 kb)
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Zhang, Q., He, C., Li, R. et al. miR-708 and miR-335-3p Inhibit the Apoptosis of Retinal Ganglion Cells Through Suppressing Autophagy. J Mol Neurosci 71, 284–292 (2021). https://doi.org/10.1007/s12031-020-01648-y
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DOI: https://doi.org/10.1007/s12031-020-01648-y