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.

References 38

[1]	张立敏,顾超,安红梅.氧化应激介导的细胞凋亡在阿尔茨海默病中的作用[J].医学综述,2021,27(9):1685-1690.ZHANG L M,GU C,AN H M.Role of oxidative stress-mediated apoptosis in Alzheimer’s disease[J].Medical Recapitulate, 2021,27(9):1685-1690.(in Chinese)
[2]	LATHE R.Hormones and the hippocampus[J].J Endocrinol, 2001,169(2):205-231.
[3]	CHIARA R B,SILVIA C,ENRICO C,et al.Corticotropin-relasing hormone (CRH) alters mitochondial morphology and function by activating the NF-κB-DRP1 axis in hippocampal neurons[J].Cell Death Dis, 2020,11(11):1004.
[4]	GAN L,YANG B Y,MEI H Y.The effect of iron dextran on the transcriptome of pig hippocampus[J].Genes Genom, 2017,39(1):1-14.
[5]	JIA J,CHEN X M,ZHU W J,et al.CART protects brain from damage through ERK activation in ischemic stroke[J].Neuropeptides, 2008,42(5-6):653-661.
[6]	JIANG H,NIU F N,ZHENG Y,et al.CART mitigates oxidative stress and DNA damage in memory deficits of APP/PS1 mice via upregulating β-amyloid metabolism-associated enzymes[J].Mol Med Rep, 2021,23(4):280.
[7]	CHANG L L,CHEN Y T,LI J,et al.Cocaine-and amphetamine-regulated transcript modulates peripheral immunity and protects against brain injury in experimental stroke[J].Brain Behav Immun, 2011,25(2):260-269.
[8]	GARLID K D,PAUCEK P,YAROV-YAROVOY V,et al.Cardioprotective effect of diazoxide and its interaction with mitochondrial ATP-Sensitive K+ channels:possible mechanism of cardioprotection[J].Circ Res, 1997,81(6):1072-1082.
[9]	QIU B,HU S D,LIU L B,et al.CART attenuates endoplasmic reticulum stress response induced by cerebral ischemia and reperfusion through upregulating BDNF synthesis and secretion[J].Biochem Biophys Res Commun, 2013,436(4):655-659.
[10]	WEI L,KAN L Y,ZENG H Y,et al.BDNF/TrkB pathway mediates the antidepressant-like role of H2S in CUMS-exposed rats by inhibition of hippocampal ER stress[J].NeuroMol Med, 2018,20(2):252-261.
[11]	PANNESE E.Morphological changes in nerve cells during normal aging[J].Brain Struct Funct, 2011,216(2):85-89.
[12]	WIEDEMANN F R,SIEMEN D,MAWRIN C,et al.The neurotrophin receptor TrkB is colocalized to mitochondrial membranes[J].Int J Biochem Cell Biol, 2006,38(4):610-620.
[13]	SISALLI M J,FELICIELLO A,NOTTE S D,et al.Nuclear-encoded NCX3 and AKAP121:Two novel modulators of mitochondrial calcium efflux in normoxic and hypoxic neurons[J].Cell Calcium, 2020,87:102193.
[14]	SZCZEPANEK K,CHEN Q,DERECKA M,et al.Mitochondrial-targeted signal transducer and activator of transcription 3 (STAT3) protects against ischemia-induced changes in the electron transport chain and the generation of reactive oxygen species[J].J Biol Chem, 2011,286(34):29610-29620.
[15]	MATTSON M P,KROEMER G.Mitochondria in cell death:novel targets for neuroprotection and cardioprotection[J].Trends Mol Med, 2003,9(5):196-205.
[16]	NOLFI-DONEGAN D,BRAGANZA A,SHIVA S.Mitochondrial electron transport chain:oxidative phosphorylation,oxidant production,and methods of measurement[J].Redox Biol, 2020,37:101674.
[17]	COBLEY J N,FIORELLO M L,BAILEY D M.13 reasons why the brain is susceptible to oxidative stress[J].Redox Biol, 2018,15:490-503.
[18]	HE Q,LI Z Y,MENG C C,et al.Parkin-dependent mitophagy is required for the inhibition of ATF4 on NLRP3 inflammasome activation in cerebral ischemia-reperfusion injury in rats[J].Cells, 2019,8(8):897.
[19]	LIU L,CAO Q,GAO W W,et al.Melatonin protects against focal cerebral ischemia-reperfusion injury in diabetic mice by ameliorating mitochondrial impairments:involvement of the Akt-SIRT3-SOD2 signaling pathway[J].Aging (Albany NY), 2021,13(12):16105-16123.
[20]	ANZELL A R,MAIZY R,PRZYKLENK K,et al.Mitochondrial quality control and disease:insights into ischemia-reperfusion injury[J].Mol Neurobiol, 2018,55(3):2547-2564.
[21]	JIN J L,LIOU A K F,SHI Y J,et al.CART treatment improves memory and synaptic structure in APP/PS1 mice[J].Sci Rep, 2015,5:10224.
[22]	HSIEH Y S,CHEN P N,YU C H,et al.Involvement of oxidative stress in the regulation of NPY/CART-mediated appetite control in amphetamine-treated rats[J].Neurotoxicology, 2015,48:131-141.
[23]	JIAO W Y,WANG Y C,KONG L H,et al.CART peptide activates the Nrf2/HO-1 antioxidant pathway and protects hippocampal neurons in a rat model of Alzheimer’s disease[J].Biochem Biophys Res Commun, 2018,501(4):1016-1022.
[24]	MAO P Z,MESHUL C K,THUILLIER P,et al.CART peptide is a potential endogenous antioxidant and preferentially localized in mitochondria[J].PLoS One, 2012,7(1):e29343.
[25]	HUSSAIN S.Measurement of nanoparticle-induced mitochondrial membrane potential alterations[M]//ZHANG Q W.Nanotoxicity.New York:Humana Press,2019,1894:123-131.
[26]	SAKTHIVEL R,MALAR D S,DEVI K P.Phytol shows anti-angiogenic activity and induces apoptosis in A549 cells by depolarizing the mitochondrial membrane potential[J].Biomed Pharmacother, 2018,105:742-752.
[27]	WANG A H,SI Z H,XUE P,et al.Tacrolimus protects hippocampal neurons of rats with status epilepticus through suppressing oxidative stress and inhibiting mitochondrial pathway of apoptosis[J].Brain Res, 2019,1715:176-181.
[28]	ZHONG J H,YU H,HUANG C,et al.Inhibition of phosphodiesterase 4 by FCPR16 protects SH-SY5Y cells against MPP+-induced decline of mitochondrial membrane potential and oxidative stress[J].Redox Biol, 2018,16:47-58.
[29]	TAN P P,ZHOU B H,ZHAO W P,et al.Mitochondria-mediated pathway regulates C2C12 cell apoptosis induced by fluoride[J].Biol Trace Elem Res, 2018,185(2):440-447.
[30]	XU T J,NIU C G,ZHANG X J,et al.β-Ecdysterone protects SH-SY5Y cells against β-amyloid-induced apoptosis via c-Jun N-terminal kinase- and Akt-associated complementary pathways[J].Lab Invest, 2018,98(4):489-499.
[31]	VENKATESAN R S,SADIQ A M M.Effect of morin-5’-sulfonic acid sodium salt on the expression of apoptosis related proteins caspase 3,bax and Bcl 2 due to the mercury induced oxidative stress in albino rats[J].Biomed Pharmacother, 2017,85:202-208.
[32]	LI L Y,JIAO Y N,JIN T Y,et al.Phenolic alkaloid oleracein E attenuates oxidative stress and neurotoxicity in AlCl3-treated mice[J].Life Sci, 2017,191:211-218.
[33]	AGCA C A,TUZCU M,GENCOGLU H,et al.Lycopene counteracts the hepatic response to 7,12-dimethylbenz[a ]anthracene by altering the expression of bax,bcl-2,caspases,and oxidative stress biomarkers[J].Pharm Biol, 2012,50(12):1513-1518.
[34]	TEJEDA G S,DÍAZ-GUERRA M.Integral characterization of defective BDNF/TrkB signalling in neurological and psychiatric disorders leads the way to new therapies[J].Int J Mol Sci, 2017,18(2):268.
[35]	LI X R,GUO C,LI Y N,et al.Ketamine administered pregnant rats impair learning and memory in offspring via the CREB pathway[J].Oncotarget, 2017,8(20):32433-32449.
[36]	LUO H Q,XIANG Y,QU X P,et al.Apelin-13 suppresses neuroinflammation against cognitive deficit in a streptozotocin-induced rat model of Alzheimer’s disease through activation of BDNF-TrkB Signaling pathway[J].Front Pharmacol, 2019,10:395.
[37]	HAN M L,LIU G H,GUO J,et al.Imipramine protects retinal ganglion cells from oxidative stress through the tyrosine kinase receptor B signaling pathway[J].Neural Regen Res, 2016,11(3):476-479.
[38]	YOO J M,LEE B D,SOK D E,et al.Neuroprotective action of N-acetyl serotonin in oxidative stress-induced apoptosis through the activation of both TrkB/CREB/BDNF pathway and Akt/Nrf2/Antioxidant enzyme in neuronal cells[J].Redox Biol, 2017,11:592-599.

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

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 38

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	ZHANG Xinting, QIU Wenyue, PANG Xiaoyue, SU Yiman, YE Jiali, HUANG Jianjia, ZHOU Shuilian, TANG Zhaoxin, WANG Rongmei, SU Rongsheng. Effect of Asiatic Acid Alleviating Myocardial Injury Caused by Lipopolysaccharide through Inhibiting Oxidative Stress and Ferroptosis in Broilers [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1787-1799.
[2]	JIANG Lijun, ZONG Yunhe, LI Yunlei, CHEN Jilan, GENG Zhaoyu, SUN Yanyan, JIN Sihua. Research Progress of Antioxidant Application in Poultry Semen Storage [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 913-923.
[3]	WANG Xin, NIE Tong, LI Aqun, MA Jun. Hesperidin Alleviates High-fat-diet Induced Hepatic Oxidative Stress in Mice via Oxidative Phosphorylation Pathway [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1302-1313.
[4]	CHEN Hong, RUAN Hongri, MA Tianwen, LI Yanan, MIAO Xue, YANG Wenyue, GAO Li, WEI Chengwei. The Mechanism of Puerarin Improving Cartilage Degeneration in PTOA Rats by Interfering with Oxidative Stress and Nrf2/HO-1 Pathway of Cartilage [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3951-3963.
[5]	MAO Peng, WANG Zhihao, LI Jianji, CUI Luying, ZHU Guoqiang, MENG Xia, DONG Junsheng, WANG Heng. Research Progress of Ferroptosis in Bacterial Infection [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2280-2287.
[6]	HAN Xiuyuan, ZHAO Liang, WANG Chuang, QI Meiyu, YAO Yuchang. Nicotinic Acid Enhances Low Temperature Preservation of Sheep Sperm by Reducing Oxidative Stress Levels [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 1979-1989.
[7]	CHEN Yongping, KOU Yuhong, JIAO Wenjing, HOU Xiaoyu, FAN Honggang. Effect of Coenzyme Q10 on LPS-induced Acute Lung Injury in Mice [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1730-1741.
[8]	XIAO Shiyu, LU Chang, MA Juan, WANG Chuang, QI Meiyu, YAO Yuchang. Effects of N-acetylcysteine [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1046-1057.
[9]	PAN Chanyuan, ZHAO Zixuan, DUAN Mingjie, JIANG Linshu, TONG Jinjin. The Mechanism of Artemisia carvifolia Alleviating Dairy Cow Oxidative Stress Predicted by Network Pharmacology [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1071-1084.
[10]	YANG Chengying, WANG Kai, HUANG Ziqing, LIN Hailan, WANG Naixiu, LI Yuhang, LIU Yanqing, LIU Yuxuan, ZHU Yan, HE Daoling, CHEN Hongyue, GAN Ling. Study on the Inhibitory Effect of Chito-oligosaccharide on Oxidative Stress inHippocampus of Piglets and Its Mechanism [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(2): 744-756.
[11]	YANG Haotian, CHEN Yongping, WANG Zhiqiang, HUANG Yuxiang, MA Zhigang, ZOU Yue, WEI Niandong, ZHANG Hong, LI Xin, DONG Jiaqiang, LÜ Mingzhe, LI Hongbin, LIU Liwei, YANG Haoxuan, ZHANG Guohua, LIU Xuesong, ZHONG Peng, SHI Heye, KOU Yuhong, CHEN Zhifeng. Dexmedetomidine Inhibits NOX4 and Alleviates Acute Stress-induced Renal Injury in Rats [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(2): 779-786.
[12]	LI Guangxing, CHEN Yang, CHEN Kaiting, WU Menglin, ZHANG Di, HUANG Xiaodan. Di (2-ethylhexyl) Phthalate Induces the Apoptosis and Necroptosis of HD11 Cells through the ROS/PTEN/PI3K/AKT Axis [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5240-5251.
[13]	LI Chunyan, ZHANG Yan, Lü Chunrong, DENG Weidong, QUAN Guobo. Research Progress on Antioxidant Mechanisms of Melatonin and Its Application in Cryopreservation of Mammalian Spermatozoa [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4468-4476.
[14]	YANG Mengran, YANG Chenglin, WU You, WANG Siqi, KONG Xiangyi, NING Can, XIAO Wenguang, FAN Hui, WU Jing, YUAN Zhihang. Study on the Mechanism of Citrinin Induced Intestinal Barrier Dysfunction in Mice [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4805-4816.
[15]	LIU Zelin, GUO Xiaoyan, LI Jiahuan, GAO Mingxing, CHENG Guofu, HU Xueying, ZHANG Wanpo, GU Changqin. Effect of the Nrf2 Pathway on Oxidative Stress Induced by Japanese Encephalitis Virus Infection of Mouse Neuroblastoma Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 317-327.