JAK-STAT通路介导gx-50针对神经元的抗氧化保护作用的研究

doi:10.3969/J.ISSN.1671-9964.2016.01.004

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Abstract

A novel compound derived from Zanthoxylum, gx-50(N-[2-(3, 4-dimethoxyphenyl)ethyl]-3-phenyl-acrylamide), has been demonstrated that it has neuroprotective effects against Alzheimer’s disease(AD)by our previous study.This study focuses on the mechanism of its antioxidant properties against AD.We measured the levels of intracellular reactive oxygen species(ROS), superoxide dismutase(SOD)activity and malondialdehyde(MDA)to determine the anti-oxidative ability of gx-50 at cellular level.We also measured gx-50’s effect on JAK-STAT signaling pathway and the relative expression levels of activated caspase-3.Data showed that gx-50 reduced the levels of reactive oxygen species(ROS)and malondialdehyde(MDA), and recovered the activity of total intracellular superoxide dismutase(SOD)in neuronal PC12 cells exposed to Aβ.After gx-50 pretreatment, the levels of p-JAK2 and p-STAT3 both increased in PC12 cells, while they were down-regulated in Aβ-treated group.In present study, we also found that gx-50 reduced the relative expression level of the activated caspase-3 in PC12 cells by activating JAK-STAT signaling pathway.Results demonstrated that gx-50 reduced amyloid-beta(Aβ)induced oxidative stress in neuron-like PC12 cells by enhancing the activation of JAK-STAT signaling pathway.It might help to protect neurons in Alzheimer's disease.

Key words： Alzheimer’s disease oxidative stress neuron gx-50 JAK-STAT pathway

Received: 29 April 2015 Published: 08 March 2016

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	Articles by authors
	Yi CHEN
	Shi SHI
	Dong-li LIANG
	Zhao-xia WANG
	Zhong-dong QIAO

Cite this article:

Yi CHEN,Shi SHI,Dong-li LIANG, et al. JAK-STAT Pathway Mediates Anti-Oxidative Effects of gx-50 on Neuron[J]. Journal of Shanghai Jiaotong University (Agricultural Sciences), 2016, 34(1): 16-23.

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http://www.qk.sjtu.edu.cn/jsjtua/EN/10.3969/J.ISSN.1671-9964.2016.01.004 OR http://www.qk.sjtu.edu.cn/jsjtua/EN/Y2016/V34/I1/16

The effect of gx-50 on the expression of intracellular ROS in neuronal PC12 cells

The values are expressed as mean ± SD, n=3-4.**P< 0.01

Effect of gx-50 on activity of total intracellular SOD in neuronal PC12 cells

Superoxide dismutase Assay Kits were used to measure the levels of activity of total intracellular SOD in PC12 cells after different treatments.The values are expressed as the mean±SD of three independent experiments.*P<0.05, **P<0.01

Effect of gx-50 on MDA production in neuronal PC12 cells

MDA Assay Kits were used to measure the levels of MDA in the medium of PC12 cells.The values are expressed as the mean ± SD of three independent experiments.**P<0.01

Effects of gx-50 on JAK-STAT signal pathway in neuronal PC12 cells

The expression levels of p-JAK2 and p-STAT3 were detected by Western blot(A), and their relative expressions compared with GAPDH were analyzed(B).The values are expressed as the mean ± SD of 3-4 independent experiments.*P< 0.05, **P< 0.01

Effect of gx-50 on JAK-STAT signaling pathway in the brain tissues of APP-Tg mice observed with immunohistochemical staining

The p-STAT3 positive neurons was stained dark brown by DAB.The nucleus were stained blue by hematoxylin

Effects of gx-50 on the relative expression levels of active casepase-3 in PC12 cells

The expression levels of active casepase-3 were detected by Western blot(A), and their relative expressions compared with procaspase-3 were analyzed(B).Data were mean ± SD error; n=3-4.** P<0.01

[1]	Praticò D.Evidence of oxidative stress in Alzheimer's disease brain and antioxidant therapy[J].Annals of the New York Academy of Sciences, 2008, 1147(1):70-78.
[2]	McLellan M E, Kajdasz S T, Hyman B T, et al.In vivo imaging of reactive oxygen species specifically associated with thioflavine S-positive amyloid plaques by multiphoton microscopy[J].The Journal of Neuroscience, 2003, 23(6):2212-2217.
[3]	Tang M, Shi S, Guo Y, et al.GSK-3/CREB pathway involved in the gx-50's effect on Alzheimer's disease[J].Neuropharmacology, 2014, 81:256-266.
[4]	Manickam M, Tulsawani R.Survival response of hippocampal neurons under low oxygen conditions induced by hippophae rhamnoides is associated with JAK/STAT signaling[J].PloS One, 2014, 9(2):e87694.
[5]	Zeng H, Chen Q, Zhao B.Genistein ameliorates beta-amyloid peptide(25-35)-induced hippocampal neuronal apoptosis[J].Free Radical biology and Medicine, 2004, 36(2):180-188.
[6]	Finkel T, Holbrook N J.Oxidants, oxidative stress and the biology of ageing[J].Nature, 2000, 408(6809):239-247.
[7]	Vanfleteren J R.Oxidative stress and ageing in Caenorhabditis elegans[J].Biochem J, 1993, 292:605-608.
[8]	Larsen P L.Aging and resistance to oxidative damage in Caenorhabditis elegans[J].Proceedings of the National Academy of Sciences, 1993, 90(19):8905-8909.
[9]	Martin G M, Austad S N, Johnson T E.Genetic analysis of ageing:role of oxidative damage and environmental stresses[J].Nature Genetics, 1996, 13(1):25-34.
[10]	Yan L J, Levine R L, Sohal R S.Oxidative damage during aging targets mitochondrial aconitase[J].Proceedings of the National Academy of Sciences, 1997, 94(21):11168-11172.
[11]	Reddy P H.Amyloid precursor protein-mediated free radicals and oxidative damage:Implications for the development and progression of Alzheimer's disease[J].Journal of Neurochemistry, 2006, 96(1):1-13.
[12]	Tang M, Wang Z, Zhou Y, et al.A novel drug candidate for Alzheimer's disease treatment:gx-50 derived from Zanthoxylum Bungeanum[J].Journal of Alzheimer's Disease, 2013, 34(1):203-213.
[13]	Guo Y, Shi S, Tang M, et al.The suppressive effects of gx-50 on Aβ-induced chemotactic migration of microglia[J].International Immunopharmacology, 2014, 19(2):283-289.
[14]	Chiba T, Yamada M, Aiso S.Targeting the JAK2/STAT3 axis in Alzheimer's disease[J].Expert Opinion on Therapeutic Targets, 2009, 13:1155-1167.
[15]	Chiba T, Yamada M, Sasabe J, et al.Amyloid-β causes memory impairment by disturbing the JAK2/STAT3 axis in hippocampal neurons[J].Molecular Psychiatry, 2008, 14(2):206-222.
[16]	Zouein F A, Duhé R J, Arany I, et al.Loss of STAT3 in mouse embryonic fibroblasts reveals its Janus-like actions on mitochondrial function and cell viability[J].Cytokine, 2014, 66(1):7-16.
[17]	Shaw S, Bencherif M, Marrero M B.Janus kinase 2, an early target of α7 nicotinic acetylcholine receptor-mediated neuroprotection against Aβ-(1-42)amyloid[J].Journal of Biological Chemistry, 2002, 277(47):44920-44924.
[18]	Szczepanek K, Lesnefsky E J, Larner A C.Multi-tasking:nuclear transcription factors with novel roles in the mitochondria[J].Trends in Cell Biology, 2012, 22(8):429-437.
[19]	Szczepanek K, Chen Q, Larner A C, et al.Cytoprotection by the modulation of mitochondrial electron transport chain:the emerging role of mitochondrial STAT3[J].Mitochondrion, 2012, 12(2):180-189.
[20]	Andreyev A Y, Kushnareva Y E, Starkov A.Mitochondrial metabolism of reactive oxygen species[J].Biochemistry(Moscow), 2005, 70(2):200-214.