[Role of PNPT1 in cardiomyocyte apoptosis induced by oxygen-glucose deprivation].

OBJECTIVE

To explore the effect of inhibiting polyribonucleotide nucleotidyl-transferase 1 (PNPT1) on oxygen-glucose deprivation (OGD)-induced apoptosis of mouse atrial myocytes.

METHODS

Cultured mouse atrial myocytes (HL-1 cells) with or without OGD were transfected with PNPT1-siRNA or a negative control siRNA (NC-siRNA group), and the cell survival rate was detected using CCK-8 assay. The expression levels of ACTB and TUBA mRNA were detected with qPCR, and the protein expression of PNPT1 was detected with Western blotting. The apoptosis rate of the treated cells was determined with flow cytometry, the mitochondrial membrane potential was detected using JC-1 kit, and the mitochondrial morphology was observed using transmission electron microscope.

RESULTS

With the extension of OGD time, the protein expression levels of PNPT1 increased progressively in the cytoplasm of HL-1 cells ( < 0.05). Transfection with PNPT1-siRNA significantly reduced PNPT1 expression in HL-1 cells ( < 0.05). Exposure to OGD significantly enhanced degradation of ACTB and TUBA mRNA ( < 0.05) and markedly increased the apoptosis rate of HL-1 cells ( < 0.05), and these changes were significantly inhibited by transfection with PNPT1-siRNA ( < 0.05), which obviously increased mitochondrial membrane potential and improved mitochondrial morphology of HL-1 cells exposed to OGD.

CONCLUSION

Inhibition of PNPT1 improves mitochondrial damage and reduces degradation of apoptotic-associated mRNAs to alleviate OGD-induced apoptosis of mouse atrial myocyte.