Inhibitory Effect of CCK-8 on Methamphetamine-Induced Apoptosis.
10.12116/j.issn.1004-5619.2021.310206
- Author:
Wu-Hua ZHANG
1
;
Ming-Long ZHANG
1
;
Wei-Wei JING
1
;
Bing XIE
1
;
Hai-Tao BI
1
;
Feng YU
1
;
Bin CONG
1
;
Chun-Ling MA
1
;
Di WEN
1
Author Information
1. Collaborative Innovation Center of Forensic Medical Molecular Identification, Hebei Key Laboratory of Forensic Medicine, College of Forensic Medicine, Hebei Medical University, Shijiazhuang 050017, China.
- Publication Type:Journal Article
- Keywords:
apoptosis;
cholecystokinin octapeptide;
forensic toxicology;
methamphetamine;
neuron;
receptor;
β-arrestin
- MeSH:
Apoptosis/physiology*;
Central Nervous System Stimulants/pharmacology*;
HEK293 Cells;
Humans;
Methamphetamine/pharmacology*;
Sincalide/pharmacology*
- From:
Journal of Forensic Medicine
2021;37(6):796-805
- CountryChina
- Language:English
-
Abstract:
OBJECTIVES:To investigate the inhibitory effect of cholecystokinin octapeptide (CCK-8) binding to cholecystokinin 2 receptor (CCK2R) on methamphetamine (METH)-induced neuronal apoptosis, and to explore the signal transduction mechanism of β-arrestin 2 in CCK-8 inhibiting METH-induced neuronal apoptosis.
METHODS:SH-SY5Y cell line was cultured, and HEK293-CCK1R and HEK293-CCK2R cell line were constructed by lentivirus transfection. Small interfering RNA (siRNA) was used to knockdown the expression of β-arrestin 2. Annexin Ⅴ-FITC/PI staining and flow cytometry were used to detect the apoptotic rate of cells, and Western blotting was used to detect the expression of apoptosis-related proteins.
RESULTS:The apoptosis of SH-SY5Y cells was induced by 1 mmol/L and 2 mmol/L METH treatment, the number of nuclear fragmentation and pyknotic cells was significantly increased, and the expression of apoptosis-related proteins Bax and cleaved caspase-3 were increased. CCK-8 pre-treatment at the dose of 0.1 mmol/L and 1 mmol/L significantly reversed METH-induced apoptosis in SH-SY5Y cells, and inhibited cell nuclear fragmentation, pyknosis and the changes of apoptosis-related proteins induced by METH. In lentivirus transfected HEK293-CCK1R and HEK293-CCK2R cells, the results revealed that CCK-8 had no significant effect on METH-induced changes of apoptosis-related proteins in HEK293-CCK1R cells, but it could inhibit the expression level of apoptosis-related proteins in HEK293-CCK2R cells induced by METH. The inhibitory effect of CCK-8 on METH-induced apoptosis was blocked by the knockdown of β-arrestin 2 expression in SH-SY5Y cells.
CONCLUSIONS:CCK-8 can bind to CCK2R and exert an inhibitory effect on METH-induced apoptosis by activating the β-arrestin 2 signal.
