Oxidized low-density lipoprotein induced mouse hippocampal HT-22 cell damage via promoting the shift from autophagy to apoptosis

CNS Neurosci Ther. 2017 Apr;23(4):341-349. doi: 10.1111/cns.12680. Epub 2017 Feb 23.

Abstract

Aims: Although oxidized low-density lipoprotein (ox-LDL) in the brain induces neuronal death, the mechanism underlying the damage effects remains largely unknown. Given that the ultimate outcome of a cell is depended on the balance between autophagy and apoptosis, this study was performed to explore whether ox-LDL induced HT-22 neuronal cell damage via autophagy impairment and apoptosis enhancement.

Methods: Flow cytometry and transmission electron microscopy (TEM) were used to evaluate changes in cell apoptosis and autophagy, respectively. The protein expression of LC3-II, p62, Bcl-2, and Bax in HT-22 cells was measured by Western bolt analysis.

Results: Our study confirmed that 100 μg/mL of ox-LDL not only promoted TH-22 cell apoptosis, characterized by elevated cell apoptosis rate and Bax protein expression, decreased Bcl-2 protein expression, and damaged cellular ultrastructures, but also impaired autophagy as indicated by the decreased LC3-II levels and the increased p62 levels. Importantly, all of these effects of ox-LDL were significantly aggravated by cotreatment with chloroquine (an inhibitor of autophagy flux). In contrast, cotreatment with rapamycin (an inducer of autophagy) remarkably reversed these effects of ox-LDL.

Conclusions: Taken together, our results indicated that ox-LDL-induced shift from autophagy to apoptosis contributes to HT-22 cell damage.

Keywords: HT-22 cell; apoptosis; autophagy; cell damage; oxidized low-density lipoprotein.

MeSH terms

  • Animals
  • Annexin A5 / metabolism
  • Antirheumatic Agents / pharmacology
  • Apoptosis / drug effects*
  • Apoptosis Regulatory Proteins / metabolism*
  • Autophagy / drug effects*
  • Cell Line, Transformed
  • Cell Survival / drug effects
  • Chloroquine / pharmacology
  • Dose-Response Relationship, Drug
  • Flow Cytometry
  • Hippocampus / cytology
  • Lipoproteins, LDL / pharmacology
  • Lipoproteins, LDL / toxicity*
  • Mice
  • Microscopy, Electron, Transmission
  • Neurons / drug effects*
  • Neurons / ultrastructure
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Sequestosome-1 Protein / metabolism
  • Sincalide / metabolism
  • Sirolimus / pharmacology
  • bcl-2-Associated X Protein / metabolism

Substances

  • Annexin A5
  • Antirheumatic Agents
  • Apoptosis Regulatory Proteins
  • Lipoproteins, LDL
  • Proto-Oncogene Proteins c-bcl-2
  • SQSTM1 protein, human
  • Sequestosome-1 Protein
  • bcl-2-Associated X Protein
  • oxidized low density lipoprotein
  • Chloroquine
  • Sincalide
  • Sirolimus