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Keywords:

  • ATP;
  • autophagy;
  • Beclin-1;
  • correlation;
  • flow cytometry

Abstract

Autophagy is usually up-regulated to provide more ATP in response to starvation or OGD (oxygen-glucose deprivation), but the relationship between autophagy and ATP, [Ca2+]i (intracellular free Ca2+ concentration) or MMP (mitochondrial membrane potential) during reoxygenation is not yet fully clear. The role of autophagy is unknown in PC12 cells subjected to 2 h OGD with different time points of reoxygenation. In the present study, we showed that Beclin-1 was up-regulated beginning at 0 h reoxygenation peaking at 24 h and lasting for 48 h. Cell viability was decreased from 0 to 48 h reoxygenation, reaching its minimum at 10 h reoxygenation. ATP was decreased from 0 to 10 h reoxygenation, reaching its minimum at 4 h reoxygenation. A significant negative correlation was observed between ATP and Beclin-1 (r = −0.61, P<0.05) at 0 h reoxygenation, but ATP was not significant related (r = 0.24, P>0.05) to Beclin-1 at 24 h reoxygenation. Besides, Nimodipine, a calcium antagonist, significantly reduced [Ca2+]i and Beclin-1, but increased MMP in OGD/R-treated cells. At 24 h reoxygenation, Beclin-1 expression reached its maximum, cell viability continued to increase, and ATP was higher than that before OGD. These results suggest that energy metabolism dysfunction can induce autophagy during OGD in PC12 cells. Increased [Ca2+]i and decreased MMP may induce autophagy during reoxygenation in PC12 cells. Autophagy may be a protective effect on PC12 cells treated with different time points of reoxygenation after 2 h OGD.