MiR-195-5p regulates oxidative stress and aerobic metabolism by directly downregulating GLS2 in high glucose-induced human lens epithelial cells.

OBJECTIVE

To investigated how miR-195-5p affects oxidative stress and modulates aerobic metabolism.

MATERIALS AND

METHODS

MiR-195-5p plus GLS2 mRNA was identified by conducting real-time quantitative polymerase chain reaction. Western blotting was conducted to determine GLS2 protein expression. Corresponding kits were used to determine the concentrations of glutamate, reduced glutathione, oxidized glutathione, a-ketoglutarate, and adenosine triphosphate. The cell counting Kit-8 assay was performed to determine viability. Flow cytometry assay was performed to measure the reactive oxygen species content. Finally, a dual-luciferase reporter assay was conducted to confirm the interaction of miR-195-5p with GLS2 mRNA in the 3'UTR.

RESULTS

In high glucose-induced SRA01/04 cells, miR-195-5p was overexpressed, and GLS2 was downregulated. When miR-195-5p was upregulated, the levels of glutamate, reduced glutathione, a-ketoglutarate, and adenosine triphosphate, along with the reduced glutathione-to-oxidized glutathione ratio decreased, whereas the reactive oxygen species levels increased. Oxidative stress was ameliorated after miR-195-5p was downregulated. MiR-195-5p adversely controls the expression of GLS2 mRNA and protein. MiR-195-5p exacerbates oxidative damage and hinders aerobic metabolism by downregulating GLS2.

CONCLUSION

Oxidative stress and aerobic metabolism in human lens epithelial cells were found to be regulated by miR-195-5p after the downregulation of GLS2.