Abstract:AIM: To assess the protective effect of adipose tissue-derived mesenchymal stem cells(ADSCs)exosomes on injured retinal ganglion cells(RGCs)by establishing an in vitro rat RGC pressure injury model.
METHODS: ADSCs were cultured, and exosomes were extracted from the supernatant and identified. Rat RGCs were divided into a control group, pressure model groups(40, 80, 120 mmHg), and exosome-treated groups under different pressures. Cell proliferation activity was assessed using the CCK-8 assay. The mRNA expression levels of brain-derived neurotrophic factor(BDNF)and Caspase-3 in RGCs were detected by qPCR, and protein levels were measured by Western Blot.
RESULTS: The CCK-8 assay showed that cell proliferation activity in the control group increased significantly at 48 h compared to 24 h(P<0.05). At 48 h, cell viability in the exosome-treated groups increased significantly compared to the 40, 80, and 120 mmHg pressure model groups(all P<0.05). qPCR results indicated that BDNF mRNA expression decreased in the 40 mmHg pressure model group without statistical significance(P>0.05), and significantly decreased in the 80 and 120 mmHg pressure model groups(all P<0.05). BDNF mRNA expression significantly increased in the 40 and 80 mmHg pressure model groups after exosome treatment(both P<0.05), and increased in the 120 mmHg pressure model group without statistical significance(P>0.05). Caspase-3 mRNA expression increased in the 40 mmHg pressure model group without statistical significance(P>0.05), and significantly increased in the 80 and 120 mmHg pressure model groups(all P<0.05). Caspase-3 mRNA expression significantly decreased in the 40 and 80 mmHg pressure model groups after exosome treatment(P<0.05), and decreased in the 120 mmHg pressure model group without statistical significance(P>0.05). Western Blot analysis showed that BDNF protein expression decreased in the 40 mmHg pressure model group without statistical significance(P>0.05), and significantly decreased in the 80 and 120 mmHg pressure model groups(all P<0.001). After exosome treatment, BDNF protein expression significantly increased compared to the pressure model groups(all P<0.05). Caspase-3 protein expression increased significantly in all pressure model groups compared to the control group(all P<0.05), and significantly decreased in all exosome-treated groups compared to the model groups(all P<0.05).
CONCLUSION: ADSCs-derived exosomes enhance cell proliferation and viability in cultured rat RGCs in vitro under different pressure-induced injuries, enhance BDNF mRNA and protein expression levels, and reduce Caspase-3 mRNA and protein expression levels, suggesting that ADSCs-derived exosomes have a protective effect on pressure-injured in cultured rat RGCs in vitro.
