Costunolide attenuates oxygen‑glucose deprivation/reperfusion‑induced mitochondrial‑mediated apoptosis in PC12 cells

Mol Med Rep. 2021 Jun;23(6):411. doi: 10.3892/mmr.2021.12050. Epub 2021 Mar 31.

Abstract

The present study investigated the effect of costunolide (CT), a compound extracted from Aucklandia lappa Decne, to attenuate oxygen‑glucose deprivation/reperfusion (OGD/R)‑induced mitochondrial‑mediated apoptosis in PC12 cells. The present study used molecular docking technology to detect the binding of CT with mitochondrial apoptotic protein targets. A model of oxygen‑glucose deprivation for 2 h and reperfusion for 24 h in PC12 cells was used to mimic cerebral ischemic injury. Cell viability and damage were measured using the Cell Counting kit‑8 and lactate dehydrogenase (LDH) cytotoxicity assay kits. Cellular apoptosis was analyzed using flow cytometry. A fluorescence microscope determined intracellular [Ca2+] and mitochondrial membrane potential. Furthermore, immunofluorescence and Western blot analyses were used to detect the expression of apoptosis‑associated proteins. CT contains binding sites with Caspase‑3, Caspase‑9 and Caspase‑7. CT markedly enhanced cell viability, inhibited LDH leakage, increased intracellular [Ca2+], stabilized the mitochondrial membrane potential, increased the expression of Bcl‑2 and inhibited the expression of Apaf‑1, Bax, cleaved‑caspase‑7, cleaved‑caspase‑9 and cleaved‑caspase‑3. CT may markedly protect PC12 cells from damage caused by OGD/R, and its mechanism is associated with blocking the calcium channel and inhibiting mitochondrial‑mediated apoptosis.

Keywords: costunolide; ischemic stroke; oxygen‑glucose deprivation/reperfusion; apoptosis; caspase; PC12 cells.

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Calcium / metabolism
  • Caspases / metabolism
  • Cell Hypoxia*
  • Glucose / deficiency*
  • L-Lactate Dehydrogenase / metabolism
  • Membrane Potential, Mitochondrial
  • Mitochondria / metabolism*
  • Neuroprotective Agents / pharmacology*
  • PC12 Cells
  • Rats
  • Sesquiterpenes / pharmacology*

Substances

  • Neuroprotective Agents
  • Sesquiterpenes
  • costunolide
  • L-Lactate Dehydrogenase
  • Caspases
  • Glucose
  • Calcium

Grants and funding

The present study was supported by the National Natural Science Foundation of Ningxia (grant no. 2020AAC02017) and the National Natural Science Foundation of China (grant no. 81660700).