外源性神经肽CART对大鼠海马神经元氧化应激的抵抗作用及机制研究

doi:10.11843/j.issn.0366-6964.2022.04.023

Abstract

Abstract: The study aimed at investigating the effect and mechanism of exogenous neuropeptide cocaine-and amphetamine-regulated transcription factor (CART) against oxidative stress in hippocampal neurons. The primary culture of hippocampal neurons was carried out in rats within 24 hours of birth. Following the purity test, 200 μmol·mL^-1 hydrogen peroxide were used to induce oxidative stress. On this basis, the effects of CART on the neuronal viability, ATP content, mitochondrial membrane potential (MMP)and the expression levels of apoptosis-related genes and trkB gene were investigated. Also the correlation between trkB gene and apoptosis-related genes was analyzed. The results showed that compared with the control group, the neuron viability and ATP content in H₂O₂ group were extremely significantly decreased(P<0.01), and the MMP was significantly decreased (P<0.05); CART with different concentrations significantly increased the neuronal viability, ATP content and MMP(P<0.05). Compared with the control group, the mRNA levels of bax and caspase-3 were extremely significantly increased in H₂O₂ group(P<0.01), and the mRNA levels of bcl-2 were extremely significantly decreased (P<0.01) and trkB were significantly decreased (P<0.05) in H₂O₂ group. Compaced with the H₂O₂ group, the mRNA levels of bcl-2 (P<0.01) were extremely significantly increased and trkB (P<0.05) were significantly increased in CART + H₂O₂ group. While the mRNA levels of bax were significantly decreased (P<0.05) and the mRNA levels of caspase-3 was extremely significantly decreased (P<0.01) in CART+H₂O₂ group compared with H₂O₂ group. The results of Western blot showed that the protein levels of bax and caspase-3 in H₂O₂ group were significantly higher than those in control group (P<0.05), and the protein levels of bcl-2 and trkB were significantly lower than those in control group (P<0.05). The protein levels of bcl-2 and trkB were significantly increased in CART+H₂O₂ group (P<0.05), while the protein levels of bax and caspase-3 were significantly decreased (P<0.05) compared with H₂O₂ group. The correlation analysis showed that, trkB gene was negatively correlated with the expression level of bax and caspase-3 genes, while positively correlated with the expression level of bcl-2 gene. The results indicate that CART can resist oxidative stress in hippocampal neurons by inhibiting apoptotic signaling pathway while increasing the expression level of trkB gene.

Key words: neuropeptide, hippocampal neurons, oxidative stress, BDNF/trkB, apoptosis signaling pathway

CLC Number:

Q539.2

YANG Chengying, LIN Hailan, HUANG Ziqing, WANG Naixiu, WANG Kai, GAN Ling. Effects and Mechanisms of Exogenous Neuropeptide CART on Oxidative Stress in Hippocampal Neurons[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1231-1240.

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Effects and Mechanisms of Exogenous Neuropeptide CART on Oxidative Stress in Hippocampal Neurons

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