Abstract
This study investigated an ethanol extract from Glycyrrhizae radix (GR), the root of Glycyrrhiza uralensis (Leguminosae), for possible neuroprotective effects on neurotoxicity induced by amyloid β protein (Aβ) (25–35) in cultured rat cortical neurons. Exposure of cultured cortical neurons to 10 μM Aβ (25–35) for 36 h induced neuronal apoptotic death. GR (10–50 μg/mL) prevented the Aβ (25–35)-induced neuronal apoptotic death, as assessed by a MTT assay and Hoechst 33342 staining. Furthermore, GR decreased the expression of Bax and active caspase-3, proapoptotic proteins, and increased Bcl-2, an antiapoptotic protein. GR also significantly inhibited Aβ (25–35)-induced elevation of the intracellular Ca2+ concentration ([Ca2+]i) and generation of reactive oxygen species (ROS) measured by fluorescent dyes. Isoliquiritigenin (1–20 μM), isolated from GR as an active component, inhibited Aβ (25–35)-induced neuronal apoptotic death, elevation of [Ca2+]i, ROS generation, and the change of apoptosis-associated proteins in cultured cortical neurons, suggesting that the neuroprotective effect of GR may be, at least partly, attributable to this compound. These results suggest that GR and isoliquiritigenin prevent Aβ (25–35)-induced neuronal apoptotic death by interfering with the increases of [Ca2+]i and ROS, and GR may have a possible therapeutic role for preventing the progression of neurodegenerative disease such as Alzheimer’s disease.
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Ahn, J., Um, M., Choi, W., Kim, S., and Ha, T., Protective effects of Glycyrrhiza uralensis Fisch. on the cognitive deficits caused by beta-amyloid peptide 25–35 in young mice. Biogerontology, 7, 239–247 (2006).
Ahn, J. Y., Kim, S., Jung, S. E., and Ha, T. Y., Effect of licorice (Glycyrrhiza uralensis Fisch) on amyloid-β-induced neurotoxicity in PC12 cells. Food Sci. Biotechnol., 19, 1391–1395 (2010).
Ban, J. Y., Jeon, S. Y., Bae, K., Song, K. S., and Seong, Y. H., Catechin and epicatechin from Smilacis chinae rhizome protect cultured rat cortical neurons against amyloid beta protein (25–35)-induced neurotoxicity through inhibition of cytosolic calcium elevation. Life Sci., 79, 2251–2259 (2006).
Behl, C., Davis, J. B., Lesley, R., and Schubert, D., Hydrogen peroxide mediates amyloid beta protein toxicity. Cell, 77, 817–827 (1994).
Block, G., Patterson, B., and Subar, A., Fruit, vegetables, and cancer prevention: a review of the epidemiological evidence. Nutr. Cancer, 18, 1–29 (1992).
Bradford, M. M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72, 248–254 (1976).
Castaldo, P., Cataldi, M., Magi, S., Lariccia, V., Arcangeli, S., and Amoroso, S., Role of the mitochondrial sodium/calcium exchanger in neuronal physiology and in the pathogenesis of neurological diseases. Prog. Neurobiol., 87, 58–79 (2009).
Cho, S. O., Ban, J. Y., Kim, J. Y., Jeong, H. Y., Lee, I. S., Song, K. S., Bae, K., and Seong, Y. H., Aralia cordata protects against amyloid beta protein (25–35)-induced neurotoxicity in cultured neurons and has antidementia activities in mice. J. Pharmacol. Sci., 111, 22–32 (2009).
Chung, W. T., Lee, S. H., Kim, J. D., Sung, N. S., Hwang, B., Lee, S. Y., Yu, C. Y., and Lee H. Y., Effect of the extracts from Glycyrrhiza uralensis Fisch on the growth characteristics of human cell lines: Anti-tumor and immune activation activities. Cytotechnology, 37, 55–64 (2001).
Costa, R. O., Ferreiro, E., Cardoso, S. M., Oliveira, C. R., and Pereira, C. M., ER stress-mediated apoptotic pathway induced by Abeta peptide requires the presence of functional mitochondria. J. Alzheimers Dis., 20, 625–636 (2010).
Cui, Y. M., Ao, M. Z., Li, W., and Yu, L. J., Effect of glabridin from Glycyrrhiza glabra on learning and memory in mice. Planta Med., 74, 377–380 (2008).
Dhingra, D., Parle, M., and Kulkarni, S. K., Memory enhancing activity of Glycyrrhiza glabra in mice. J. Ethnopharmacol., 91, 361–365 (2004).
Golde, T. E., Estus, S., Younkin, L. H., Selkoe, D. J., and Younkin, S. G., Processing of the amyloid protein precursor to potentially amyloidogenic derivatives. Science, 255, 728–730 (1992).
Hockenbery, D. M., Oltvai, Z. N., Yin, X. M., Milliman, C. L., and Korsmeyer, S. J., Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell, 75, 241–251 (1993).
Hong, D. J., Pei, A. L., Sun, F. Y., and Zhu, C. Q., Aberrant neuronal expression of mitotic protein, tau and Bax in the rat brain after injection of Abeta(25–35) into the amygdala. Sheng Li Xue Bao, 55, 142–146 (2003).
Hwang, I. K., Lim, S. S., Choi, K. H., Yoo, K. Y., Shin, H. K., Kim, E. J., Yoon-Park, J. H., Kang, T. C., Kim, Y. S., Kwon, D. Y., Kim, D. W., Moon, W. K., and Won, M. H., Neuroprotective effects of roasted licorice, not raw form, on neuronal injury in gerbil hippocampus after transient forebrain ischemia. Acta Pharmacol. Sin., 27, 959–965 (2006).
Jeong, H. Y., Kim, J. Y., Lee, H. K., Ha do, T., Song, K. S., Bae, K., and Seong, Y. H., Leaf and stem of Vitis amurensis and its active components protect against amyloid β protein (25–35)-induced neurotoxicity. Arch. Pharm. Res., 33, 1655–1664 (2010)
Keli, S. O., Hertog, M. G., Feskens, E. J., and Kromhout, D., Dietary flavonoids, antioxidant vitamins, and incidence of stroke: the Zutphen study. Arch. Intern. Med., 156, 637–642 (1996).
Kim, H. S., Lee, J. H., Lee, J. P., Kim, E. M., Chang, K. A., Park, C. H., Jeong, S. J., Wittendorp, M. C., Seo, J. H., Choi, S. H., and Suh, Y. H., Amyloid beta peptide induces cytochrome C release from isolated mitochondria. Neuroreport, 13, 1989–1993 (2002).
Kim, J. Y., Park, S. J., Yun, K. J., Cho, Y. W., Park, H. J., and Lee, K. T., Isoliquiritigenin isolated from the roots of Glycyrrhiza uralensis inhibits LPS-induced iNOS and COX-2 expression via the attenuation of NF-kappaB in RAW 264.7 macrophages. Eur. J. Pharmacol., 584, 175–184 (2008).
Kumar, S., Sharma, A., Madan, B., Singhal, V., and Ghosh, B., Isoliquiritigenin inhibits IkappaB kinase activity and ROS generation to block TNF-alpha induced expression of cell adhesion molecules on human endothelial cells. Biochem. Pharmacol., 73, 1602–1612 (2007).
Kuperstein, F. and Yavin, E., Pro-apoptotic signaling in neuronal cells following iron and amyloid beta peptide neurotoxicity. J. Neurochem., 86, 114–125 (2003).
Mailly, F., Marin, P., Israël, M., Glowinski, J., and Prémont, J., Increase in external glutamate and NMDA receptor activation contribute to H2O2-induced neuronal apoptosis. J. Neurochem., 73, 1181–1188 (1999).
Mattson, M. P., Cheng, B., Davis, D., Bryant, K., Lieberburg, I., and Rydel, R. E., beta-Amyloid peptides destabilize calcium homeostasis and render human cortical neurons vulnerable to excitotoxicity. J. Neurosci., 12, 376–389 (1992).
Missiaen, L., Callewaert, G., Parys, J. B., Wuytack, F., Raeymaekers, L., Droogmans, G., Nilius, B., Eggermont, J., and De Smedt, H., Intracellular calcium: physiology and physiopathology. Verh. K. Acad. Geneeskd. Belg., 62, 471–499 (2000).
Ohyagi, Y., Yamada, T., Nishioka, K., Clarke, N. J., Tomlinson, A. J., Naylor, S., Nakabeppu, Y., Kira, J., and Younkin, S. G., Selective increase in cellular A beta 42 is related to apoptosis but not necrosis. Neuroreport, 11, 167–171 (2000).
Pereira, C., Santos, M. S., and Oliveira, C., Mitochondrial function impairment induced by amyloid beta-peptide on PC12 cells. Neuroreport, 9, 1749–1755 (1998).
Pike, C. J., Walencewicz-Wasserman, A. J., Kosmoski, J., Cribbs, D. H., Glabe, C. G., and Cotman, C. W., Structureactivity analyses of beta-amyloid peptides: contributions of the beta 25-35 region to aggregation and neurotoxicity. J. Neurochem., 64, 253–265 (1995).
Shin, E. M., Zhou, H. Y., Guo, L. Y., Kim, J. A., and Lee, S. H., Merfort I, Kang SS, Kim HS, Kim S, Kim YS. Antiinflammatory effects of glycyrol isolated from Glycyrrhiza uralensis in LPS-stimulated RAW264.7 macrophages. Int. Immunopharmacol., 8, 1524–1532 (2008).
Shin, Y. W., Bae, E. A., Lee, B., Lee, S. H., Kim, J. A., Kim, Y. S., and Kim, D. H., In vitro and in vivo antiallergic effects of Glycyrrhiza glabra and its components. Planta Med., 73, 257–261 (2007).
Sun, Y. X., Tang, Y., Wu, A. L., Liu, T., Dai, X. L., Zheng, Q. S., and Wang, Z. B., Neuroprotective effect of liquiritin against focal cerebral ischemia/reperfusion in mice via its antioxidant and antiapoptosis properties. J. Asian Nat. Prod. Res., 12, 1051–1060 (2010).
Tang, S. Y., Whiteman, M., Peng, Z. F., Jenner, A., Yong, E. L., and Halliwell, B., Characterization of antioxidant and antiglycation properties and isolation of active ingredients from traditional chinese medicines. Free Radic. Biol. Med., 36, 1575–1587 (2004).
Tijburg, L. B., Mattern, T., Folts, J. D., Weisgerber, U. M., and Katan, M. B., Tea flavonoids and cardiovascular disease: a review. Crit. Rev. Food Sci. Nutr., 37, 771–785 (1997).
Ueda, K., Shinohara, S., Yagami, T., Asakura, K., and Kawasaki, K., Amyloid beta protein potentiates Ca2+ influx through L-type voltage-sensitive Ca2+ channels: a possible involvement of free radicals. J. Neurochem., 68, 265–271 (1997).
Vaisid, T., Barnoy, S., and Kosower, N. S., Calpain activates caspase-8 in neuron-like differentiated PC12 cells via the amyloid-beta-peptide and CD95 pathways. Int. J. Biochem. Cell Biol., 41, 2450–2458 (2009).
Wang, W., Hu, X., Zhao, Z., Liu, P., Hu, Y., Zhou, J., Zhou, D., Wang, Z., Guo, D., and Guo, H., Antidepressant-like effects of liquiritin and isoliquiritin from Glycyrrhiza uralensis in the forced swimming test and tail suspension test in mice. Prog. Neuropsychopharmacol. Biol. Psychiatry, 32, 1179–1184 (2008).
Yankner, B. A., Mechanisms of neuronal degeneration in Alzheimer’s disease. Neuron, 16, 921–932 (1996).
Yu, S. M. and Kuo, S. C., Vasorelaxant effect of isoliquiritigenin, a novel soluble guanylate cyclase activator, in rat aorta. Br. J. Pharmacol., 114, 1587–1594 (1995).
Zhan, C. and Yang, J., Protective effects of isoliquiritigenin in transient middle cerebral artery occlusion-induced focal cerebral ischemia in rats. Pharmacol. Res., 53, 303–309 (2006).
Zheng, Z.-P., Cheng, K.-W., Chao, J., Wu, J., and Wang, M., Tyrosinase inhibitors from paper mulberry (Broussonetia papyrifera). Food Chem., 106, 529–535 (2008).
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Lee, H.K., Yang, EJ., Kim, J.Y. et al. Inhibitory effects of glycyrrhizae radix and its active component, isoliquiritigenin, on Aβ(25–35)-induced neurotoxicity in cultured rat cortical neurons. Arch. Pharm. Res. 35, 897–904 (2012). https://doi.org/10.1007/s12272-012-0515-y
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DOI: https://doi.org/10.1007/s12272-012-0515-y