Open AccessCostunolide attenuates oxygen‑glucose deprivation/reperfusion‑induced mitochondrial‑mediated apoptosis in PC12 cells
Author(s) -
Meng Li,
Huixia Ma,
Jinni Meng,
Tingting Li,
Yu Zhu,
Qipeng Zhao
Publication year - 2021
Publication title -
molecular medicine reports
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 0.727
H-Index - 56
eISSN - 1791-3004
pISSN - 1791-2997
DOI - 10.3892/mmr.2021.12050
Subject(s) - apoptosis , viability assay , microbiology and biotechnology , intracellular , biology , western blot , mitochondrion , mitochondrial apoptosis induced channel , cell , lactate dehydrogenase , flow cytometry , inner mitochondrial membrane , biochemistry , enzyme , gene
The present study investigated the effect of costunolide (CT), a compound extracted from A ucklandia 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 [Ca 2+ ] 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 [Ca 2+ ], 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.