Effect of cocaine-and amphetamine-regulated transcription peptide on synaptic formation in cultured cortical neurons subjected to oxygen-glucose deprivation
10.3760/cma.j.issn.1673-4165.2020.06.006
- VernacularTitle:可卡因－苯丙胺调节转录肽对氧葡萄糖剥夺的培养皮质神经元突触形成的影响
- Author:
Luna WANG
1
;
Xiang CAO
;
Zhi ZHANG
;
Jian QIAN
;
Dujuan SHA
Author Information
1. 南京大学医学院附属鼓楼医院全科医学科 210008
- From:
International Journal of Cerebrovascular Diseases
2020;28(6):433-439
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of cocaine- and amphetamine-regulated transcript peptide (CART) on the synapse structure of mice cortical neuron subjected to oxygen-glucose deprivation (OGD).Methods:Primary neurons of the embryonic cerebral cortex obtained from healthy and clean Kunming mice at gestational age of 16-17 d were cultured. They were divided into control group, CART group, OGD group, and OGD+ CART group. 0.4 nmol/L CART 55-102 was added and cultured for 12 h after OGD treatment in the OGD+ CART group; the CART group was given the same dose of CART 55-102. The neuronal mortality was measured by the flow cytometry. The changes of synaptic structure were observed by immunofluorescence analysis, and the axon length and synapsin Ⅰ positive area were quantitatively analyzed. Real-time fluorescent quantitative polymerase chain reaction and Western blot analysis were used to identify the brain-derived neurotrophic factor (BDNF) mRNA and protein expression. Results:Compared with the control group, the mortality of neurons in the OGD group was significantly increased, the neuronal synapse growth was significantly inhibited, the positive area of synapsin Ⅰ was significantly reduced, and the expression levels of BDNF mRNA and protein were significantly down-regulated (all P<0.05). Compared with the OGD group, adding CART 55-102 significantly reduced the mortality of OGD neurons ( P<0.05), reversed the inhibitory effect of OGD on neuronal synapse growth, significantly increased the length of neuron axons and the positive area of synapsin Ⅰ (all P<0.05), and significantly up-regulated BDNF mRNA and protein expression levels (all P<0.05). Conclusion:CART can protect the synaptic structure of mice cortical neuron subjected to OGD, and its mechanism may be related to the up-regulation of BDNF expression.
