Abstract
Our previous studies demonstrated that the multifunctional agent TBN, a derivative of tetramethylpyrazine armed with a nitrone moiety, displayed high therapeutic efficacy in experimental ischemic stroke models. However, its molecular mechanisms of action underlying the neuroprotective effect need further exploration. In the present study, we found that TBN had significant activities scavenging free radicals such as ·OH, O ·−2 and ONOO−, inhibiting Ca2+ overload, maintaining mitochondrial function and preventing neuronal damage in primary cortical cultures. Further, TBN was effective in reducing brain infarction and ameliorating impairment of behavioral functions in the permanent middle cerebral artery occlusion (p-MCAo) rat model. TBN down-regulated the expression of pro-apoptotic factors Bax, while up-regulated the expression of anti-apoptotic factor Bcl-2 and increased the expression of pro-survival factors including p-Akt and p-GSK3β in the peri-infarct cortex of p-MCAo rats. In addition, LY-294002 (a PI3K inhibitor) and MK2206 (an Akt inhibitor) significantly blocked the protective effect of TBN against OGD-induced death of cortical neurons. Taken together, the multifunctional mechanisms including scavenging free radicals, blocking calcium overload, maintaining mitochondrial function and activating the PI3K/Akt/p-GSK3β cell survival pathway were possibly involved in the neuroprotective effects of TBN, making it a promising clinical candidate for the treatment of ischemic stroke.
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References
Allen, C. L., & Bayraktutan, U. (2009). Oxidative stress and its role in the pathogenesis of ischaemic stroke. International Journal of Stroke, 4(6), 461–470. https://doi.org/10.1111/j.1747-4949.2009.00387.x.
Aruoma, O. I., Kaur, H., & Halliwell, B. (1991). Oxygen free radicals and human diseases. Journal of the Royal Society of Health, 111(5), 172–177.
Beckman, J. S. (1996). Oxidative damage and tyrosine nitration from peroxynitrite. Chemical Research in Toxicology, 9(5), 836–844. https://doi.org/10.1021/tx9501445.
Chamorro, A., Dirnagl, U., Urra, X., & Planas, A. M. (2016). Neuroprotection in acute stroke: Targeting excitotoxicity, oxidative and nitrosative stress, and inflammation. The Lancet Neurology, 15(8), 869–881. https://doi.org/10.1016/S1474-4422(16)00114-9.
Chen, J., Li, Y., Wang, L., Zhang, Z., Lu, D., Lu, M., et al. (2001). Therapeutic benefit of intravenous administration of bone marrow stromal cells after cerebral ischemia in rats. Stroke, 32(4), 1005–1011.
Domin, H., Przykaza, L., Jantas, D., Kozniewska, E., Boguszewski, P. M., & Smialowska, M. (2016). Neuroprotective potential of the group III mGlu receptor agonist ACPT-I in animal models of ischemic stroke: In vitro and in vivo studies. Neuropharmacology, 102, 276–294. https://doi.org/10.1016/j.neuropharm.2015.11.025.
Feuerstein, G. Z., Zaleska, M. M., Krams, M., Wang, X., Day, M., Rutkowski, J. L., et al. (2008). Missing steps in the STAIR case: A Translational Medicine perspective on the development of NXY-059 for treatment of acute ischemic stroke. Journal of Cerebral Blood Flow and Metabolism, 28(1), 217–219. https://doi.org/10.1038/sj.jcbfm.9600516.
Fisher, M., Feuerstein, G., Howells, D. W., Hurn, P. D., Kent, T. A., Savitz, S. I., et al. (2009). Update of the stroke therapy academic industry roundtable preclinical recommendations. Stroke, 40(6), 2244–2250. https://doi.org/10.1161/strokeaha.108.541128.
Galluzzi, L., Blomgren, K., & Kroemer, G. (2009). Mitochondrial membrane permeabilization in neuronal injury. Nature Reviews Neuroscience, 10(7), 481–494. https://doi.org/10.1038/nrn2665.
Ginsberg, M. D. (2007). Life after cerovive: A personal perspective on ischemic neuroprotection in the post-NXY-059 era. Stroke, 38(6), 1967–1972. https://doi.org/10.1161/STROKEAHA.106.479170.
Guo, B., Xu, D., Duan, H., Du, J., Zhang, Z., Lee, S. M., et al. (2014). Therapeutic effects of multifunctional tetramethylpyrazine nitrone on models of Parkinson’s disease in vitro and in vivo. Biological & Pharmaceutical Bulletin, 37(2), 274–285.
Hacke, W., Kaste, M., Bluhmki, E., Brozman, M., Davalos, A., Guidetti, D., et al. (2008). Thrombolysis with alteplase 3 to 4.5 hours after acute ischemic stroke. New England Journal of Medicine, 359(13), 1317–1329. https://doi.org/10.1056/NEJMoa0804656.
Hur, E. M., & Zhou, F. Q. (2010). GSK3 signalling in neural development. Nature Reviews Neuroscience, 11(8), 539–551. https://doi.org/10.1038/nrn2870.
Kornhuber, J., & Quack, G. (1995). Cerebrospinal fluid and serum concentrations of the N-methyl-d-aspartate (NMDA) receptor antagonist memantine in man. Neuroscience Letters, 195(2), 137–139.
Kuroda, S., Tsuchidate, R., Smith, M. L., Maples, K. R., & Siesjo, B. K. (1999). Neuroprotective effects of a novel nitrone, NXY-059, after transient focal cerebral ischemia in the rat. Journal of Cerebral Blood Flow and Metabolism, 19(7), 778–787. https://doi.org/10.1097/00004647-199907000-00008.
Lai, T. W., Zhang, S., & Wang, Y. T. (2014). Excitotoxicity and stroke: Identifying novel targets for neuroprotection. Progress in Neurobiology, 115, 157–188. https://doi.org/10.1016/j.pneurobio.2013.11.006.
Lees, K. R., Zivin, J. A., Ashwood, T., Davalos, A., Davis, S. M., Diener, H. C., et al. (2006). NXY-059 for acute ischemic stroke. New England Journal of Medicine, 354(6), 588–600. https://doi.org/10.1056/NEJMoa052980.
Lin, Y., Zhang, J. C., Fu, J., Chen, F., Wang, J., Wu, Z. L., et al. (2013). Hyperforin attenuates brain damage induced by transient middle cerebral artery occlusion (MCAO) in rats via inhibition of TRPC6 channels degradation. Journal of Cerebral Blood Flow and Metabolism, 33(2), 253–262. https://doi.org/10.1038/jcbfm.2012.164.
Loncarevic-Vasiljkovic, N., Milanovic, D., Pesic, V., Tesic, V., Brkic, M., Lazic, D., et al. (2016). Dietary restriction suppresses apoptotic cell death, promotes Bcl-2 and Bcl-xl mRNA expression and increases the Bcl-2/Bax protein ratio in the rat cortex after cortical injury. Neurochemistry International, 96, 69–76. https://doi.org/10.1016/j.neuint.2016.02.017.
Long, C., Chen, M. F., Sarwinski, S. J., Chen, P. Y., Si, M., Hoffer, B. J., et al. (2006). Monoamine uptake inhibitors block alpha7-nAChR-mediated cerebral nitrergic neurogenic vasodilation. American Journal of Physiology Heart and Circulatory Physiology, 291(1), H202–H209. https://doi.org/10.1152/ajpheart.01192.2005.
Margaill, I., Plotkine, M., & Lerouet, D. (2005). Antioxidant strategies in the treatment of stroke. Free Radical Biology and Medicine, 39(4), 429–443. https://doi.org/10.1016/j.freeradbiomed.2005.05.003.
McCracken, E., Valeriani, V., Simpson, C., Jover, T., McCulloch, J., & Dewar, D. (2000). The lipid peroxidation by-product 4-hydroxynonenal is toxic to axons and oligodendrocytes. Journal of Cerebral Blood Flow and Metabolism, 20(11), 1529–1536. https://doi.org/10.1097/00004647-200011000-00002.
Minnerup, J., Wersching, H., Schilling, M., & Schabitz, W. R. (2014). Analysis of early phase and subsequent phase III stroke studies of neuroprotectants: Outcomes and predictors for success. Experimental & Translational Stroke Medicine, 6(1), 2. https://doi.org/10.1186/2040-7378-6-2.
Moskowitz, M. A., Lo, E. H., & Iadecola, C. (2010). The science of stroke: Mechanisms in search of treatments. Neuron, 67(2), 181–198. https://doi.org/10.1016/j.neuron.2010.07.002.
Oyama, Y., Carpenter, D. O., Chikahisa, L., & Okazaki, E. (1996). Flow-cytometric estimation on glutamate- and kainate-induced increases in intracellular Ca2+ of brain neurons: A technical aspect. Brain Research, 728(1), 121–124.
Park, S. Y., Yoo, S. S., Uh, J. H., Eun, J. B., Lee, H. C., Kim, Y. J., et al. (2007). Evaluation of lipid oxidation and oxidative products as affected by pork meat cut, packaging method, and storage time during frozen storage (−10 °C). Journal of Food Science, 72(2), C114–C119. https://doi.org/10.1111/j.1750-3841.2006.00265.x.
Raz, L., Zhang, Q. G., Zhou, C. F., Han, D., Gulati, P., Yang, L. C., et al. (2010). Role of Rac1 GTPase in NADPH oxidase activation and cognitive impairment following cerebral ischemia in the rat. PLoS ONE, 5(9), e12606. https://doi.org/10.1371/journal.pone.0012606.
Rhee, S. G. (1999). Redox signaling: Hydrogen peroxide as intracellular messenger. Experimental & Molecular Medicine, 31(2), 53–59. https://doi.org/10.1038/emm.1999.9.
Sairanen, T., Karjalainen-Lindsberg, M. L., Paetau, A., Ijas, P., & Lindsberg, P. J. (2006). Apoptosis dominant in the periinfarct area of human ischaemic stroke—A possible target of antiapoptotic treatments. Brain, 129(Pt 1), 189–199. https://doi.org/10.1093/brain/awh645.
Savitz, S. I., & Fisher, M. (2007). Future of neuroprotection for acute stroke: In the aftermath of the SAINT trials. Annals of Neurology, 61(5), 396–402. https://doi.org/10.1002/ana.21127.
Seira, O., & Del Rio, J. A. (2014). Glycogen synthase kinase 3 beta (GSK3beta) at the tip of neuronal development and regeneration. Molecular Neurobiology, 49(2), 931–944. https://doi.org/10.1007/s12035-013-8571-y.
Shuaib, A., Lees, K. R., Lyden, P., Grotta, J., Davalos, A., Davis, S. M., et al. (2007). NXY-059 for the treatment of acute ischemic stroke. New England Journal of Medicine, 357(6), 562–571. https://doi.org/10.1056/NEJMoa070240.
Stone, J. R., & Yang, S. (2006). Hydrogen peroxide: A signaling messenger. Antioxidants & Redox Signaling, 8(3–4), 243–270. https://doi.org/10.1089/ars.2006.8.243.
Sun, Y., Jiang, J., Zhang, Z., Yu, P., Wang, L., Xu, C., et al. (2008). Antioxidative and thrombolytic TMP nitrone for treatment of ischemic stroke. Bioorganic & Medicinal Chemistry, 16(19), 8868–8874. https://doi.org/10.1016/j.bmc.2008.08.075.
Sun, Y., Yu, P., Zhang, G., Wang, L., Zhong, H., Zhai, Z., et al. (2012). Therapeutic effects of tetramethylpyrazine nitrone in rat ischemic stroke models. Journal of Neuroscience Research, 90(8), 1662–1669. https://doi.org/10.1002/jnr.23034.
Tian, X., Zhang, L., Wang, J., Dai, J., Shen, S., Yang, L., et al. (2013). The protective effect of hyperbaric oxygen and Ginkgo biloba extract on Abeta25-35-induced oxidative stress and neuronal apoptosis in rats. Behavioural Brain Research, 242, 1–8. https://doi.org/10.1016/j.bbr.2012.12.026.
Ueno, Y., Chopp, M., Zhang, L., Buller, B., Liu, Z., Lehman, N. L., et al. (2012). Axonal outgrowth and dendritic plasticity in the cortical peri-infarct area after experimental stroke. Stroke, 43(8), 2221–2228. https://doi.org/10.1161/STROKEAHA.111.646224.
Visavadiya, N. P., McEwen, M. L., Pandya, J. D., Sullivan, P. G., Gwag, B. J., & Springer, J. E. (2013). Antioxidant properties of Neu 2000 on mitochondrial free radicals and oxidative damage. Toxicology In Vitro, 27(2), 788–797. https://doi.org/10.1016/j.tiv.2012.12.011.
Vogiatzi, T., Xilouri, M., Vekrellis, K., & Stefanis, L. (2008). Wild type alpha-synuclein is degraded by chaperone-mediated autophagy and macroautophagy in neuronal cells. The Journal of Biological Chemistry, 283(35), 23542–23556. https://doi.org/10.1074/jbc.M801992200.
Writing Group Members, Mozaffarian, D., Benjamin, E. J., Go, A. S., Arnett, D. K., Blaha, M. J., et al. (2016). Heart disease and stroke statistics-2016 update: A report from the American Heart Association. Circulation, 133(4), e38–360. https://doi.org/10.1161/cir.0000000000000350.
Ye, R., Yang, Q., Kong, X., Li, N., Zhang, Y., Han, J., et al. (2012). Sevoflurane preconditioning improves mitochondrial function and long-term neurologic sequelae after transient cerebral ischemia: Role of mitochondrial permeability transition. Critical Care Medicine, 40(9), 2685–2693. https://doi.org/10.1097/CCM.0b013e318258fb90.
Zhang, T., Gu, J., Wu, L., Li, N., Sun, Y., Yu, P., et al. (2017). Neuroprotective and axonal outgrowth-promoting effects of tetramethylpyrazine nitrone in chronic cerebral hypoperfusion rats and primary hippocampal neurons exposed to hypoxia. Neuropharmacology, 118, 137–147. https://doi.org/10.1016/j.neuropharm.2017.03.022.
Zhang, Z., Zhang, G., Sun, Y., Szeto, S. S., Law, H. C., Quan, Q., et al. (2016a). Tetramethylpyrazine nitrone, a multifunctional neuroprotective agent for ischemic stroke therapy. Scientific Reports, 6, 37148. https://doi.org/10.1038/srep37148.
Zhang, G., Zhang, F., Zhang, T., Gu, J., Li, C., Sun, Y., et al. (2016b). Tetramethylpyrazine nitrone improves neurobehavioral functions and confers neuroprotection on rats with traumatic brain injury. Neurochemical Research, 41(11), 2948–2957. https://doi.org/10.1007/s11064-016-2013-y.
Zhao, Y., Liu, Y., & Chen, K. (2016). Mechanisms and clinical application of tetramethylpyrazine (an interesting natural compound isolated from Ligusticum wallichii): Current status and perspective. Oxidative Medicine and Cellular Longevity, 2016, 2124638. https://doi.org/10.1155/2016/2124638.
Zhao, H., Tao, Z., Wang, R., Liu, P., Yan, F., Li, J., et al. (2014). MicroRNA-23a-3p attenuates oxidative stress injury in a mouse model of focal cerebral ischemia-reperfusion. Brain Research, 1592, 65–72. https://doi.org/10.1016/j.brainres.2014.09.055.
Zhou, X., Zhu, L., Wang, L., Guo, B., Zhang, G., Sun, Y., et al. (2015). Protective effect of edaravone in primary cerebellar granule neurons against iodoacetic acid-induced cell injury. Oxidative Medicine and Cellular Longevity, 2015, 606981. https://doi.org/10.1155/2015/606981.
Acknowledgements
This study was supported in part by the Scientific Projects of Guangdong Province (2013A022100030, 2015A020211019, 2015B020232011, 2016A030313107); the National Science Foundation of China (81502908); the Science and Technology Program of Guangzhou (201704020181); and the Fundamental Research Funds for the Central Universities (21617469).
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Zhang, G., Zhang, T., Wu, L. et al. Neuroprotective Effect and Mechanism of Action of Tetramethylpyrazine Nitrone for Ischemic Stroke Therapy. Neuromol Med 20, 97–111 (2018). https://doi.org/10.1007/s12017-018-8478-x
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DOI: https://doi.org/10.1007/s12017-018-8478-x