Muscone inhibits opening of mPTP to alleviate OGD/R-induced injury of HT22 cells.
10.19540/j.cnki.cjcmm.20230712.703
- Author:
Ping HUANG
1
;
Mei-Ling YUAN
1
;
Lei WANG
1
;
Yu-Ang CHEN
2
;
Ning WANG
1
;
Si-Peng WU
3
Author Information
1. Anhui Province Key Laboratory of Research & Development of Chinese Medicine, Anhui University of Chinese Medicine Hefei 230012, China Anhui Provincial Key Laboratory of Traditional Chinese Medicine Compounds Hefei 230012, China Institute for Pharmacodynamics and Safety Evaluation of Chinese Medicine, Anhui Academy of Traditional Chinese Medicine Hefei 230012, China.
2. Anhui Province Key Laboratory of Research & Development of Chinese Medicine, Anhui University of Chinese Medicine Hefei 230012, China Anhui Provincial Key Laboratory of Traditional Chinese Medicine Compounds Hefei 230012, China.
3. Anhui Province Key Laboratory of Research & Development of Chinese Medicine, Anhui University of Chinese Medicine Hefei 230012, China State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen University Xiamen 361005, China.
- Publication Type:Journal Article
- Keywords:
OGD/R;
apoptosis;
mitochondrial dysfunction;
mitochondrial permeability transition pore;
muscone
- MeSH:
Mice;
Animals;
Reactive Oxygen Species/metabolism*;
Molecular Docking Simulation;
Apoptosis;
Oxygen;
Adenosine Triphosphate/pharmacology*;
Mitogen-Activated Protein Kinase Kinases/pharmacology*;
Glucose/metabolism*
- From:
China Journal of Chinese Materia Medica
2023;48(22):6154-6163
- CountryChina
- Language:Chinese
-
Abstract:
This study aims to investigate the mechanism of muscone in inhibiting the opening of mitochondrial permeability transition pore(mPTP) to alleviate the oxygen and glucose deprivation/reoxygenation(OGD/R)-induced injury of mouse hippocampal neurons(HT22). An in vitro model of HT22 cells injured by OGD/R was established. CCK-8 assay was employed to examine the viability of HT22 cells, fluorescence microscopy to measure the mitochondrial membrane potential, the content of reactive oxygen species(ROS), and the opening of mPTP in HT22 cells. Enzyme-linked immunosorbent assay was employed to determine the level of ATP and the content of cytochrome C(Cyt C) in mitochondria of HT22 cells. Flow cytometry was employed to determine the Ca~(2+) content and apoptosis of HT22 cells. The expression of Bcl-2(B-cell lymphoma-2) and Bcl-2-associated X protein(Bax) was measured by Western blot. Molecular docking and Western blot were employed to examine the binding between muscone and methyl ethyl ketone(MEK) after pronase hydrolysis of HT22 cell proteins. After the HT22 cells were treated with U0126, an inhibitor of MEK, the expression levels of MEK, p-ERK, and CypD were measured by Western blot. The results showed that compared with the OGD/R model group, muscone significantly increased the viability, mitochondrial ATP activity, and mitochondrial membrane potential, lowered the levels of ROS, Cyt C, and Ca~(2+), and reduced mPTP opening to inhibit the apoptosis of HT22 cells. In addition, muscone up-regulated the expression of MEK, p-ERK, and down-regulated that of CypD. Molecular docking showed strong binding activity between muscone and MEK. In conclusion, muscone inhibits the opening of mPTP to inhibit apoptosis, thus exerting a protective effect on OGD/R-injured HT22 cells, which is associated with the activation of MEK/ERK/CypD signaling pathway.
