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

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.