Abstract
Accumulated evidence has shown that excessive reactive oxygen species (ROS) have been implicated in neuronal cell death related with various chronic neurodegenerative disorders. This study was designed to explore neuroprotective effects of ethyl acetate extract of Arctium lappa L. roots (EAL) on hydrogen peroxide (H2O2)-induced cell injury in human SH-SY5Y neuroblastoma cells. The cell viability was significantly decreased after exposure to 200 μM H2O2, whereas pretreatment with different concentrations of EAL attenuated the H2O2-induced cytotoxicity. Hoechst 33342 staining indicated that EAL reversed nuclear condensation in H2O2-treated cells. Meanwhile, TUNEL assay with DAPI staining showed that EAL attenuated apoptosis was induced by H2O2. Pretreatment with EAL also markedly elevated activities of antioxidant enzyme (GSH-Px and SOD), reduced lipid peroxidation (MDA) production, prevented ROS formation, and the decrease of mitochondrial membrane potential. In addition, EAL showed strong radical scavenging ability in 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) assays. Furthermore, EAL inhibited H2O2-induced apoptosis by increases in the Bcl-2/Bax ratio, decreases in cytochrome c release, and attenuation of caspase-3, caspase-9 activities, and expressions. These findings suggest that EAL may be regarded as a potential antioxidant agent and possess potent neuroprotective activity against H2O2-induced injury.
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References
Azmi NH, Ismail N, Imam MU, Ismail M (2013) Ethyl acetate extract of germinated brown rice attenuates hydrogen peroxide-induced oxidative stress in human SH-SY5Y neuroblastoma cells: role of anti-apoptotic, pro-survival and antioxidant genes. BMC Complement Altern Med 13:177. doi:10.1186/1472-6882-13-177
Beal MF (2005) Mitochondria take center stage in aging and neurodegeneration. Ann Neurol 58:495–505. doi:10.1002/ana.20624
Choi DJ, Kim SL, Choi JW, Park YI (2014) Neuroprotective effects of corn silk maysin via inhibition of H2O2-induced apoptotic cell death in SK-N-MC cells. Life Sci 109:57–64. doi:10.1016/j.lfs.2014.05.020
de Almeida ABA, Luiz-Ferreira A, Cola M, Magri LD, Batista LM, de Paiva JA, Trigo JR, Souza-Brito ARM (2012) Anti-ulcerogenic mechanisms of the sesquiterpene lactone onopordopicrin-enriched fraction from Arctium lappa L. (Asteraceae): role of somatostatin, gastrin, and endogenous sulfhydryls and nitric oxide. J Med Food 15:378–383. doi:10.1089/jmf.2011.0025
Dumont M, Beal MF (2011) Neuroprotective strategies involving ROS in Alzheimer disease. Free Radical Bio Med 51:1014–1026. doi:10.1016/j.freeradbiomed.2010.11.026
Gentil M, Pereira JV, Sousa YTCS, Pietro R, Neto MDS, Vansan LP, de Castro França S (2006) In vitro evaluation of the antibacterial activity of Arctium lappa as a phytotherapeutic agent used in intracanal dressings. Phytother Res 20:184–186. doi:10.1002/ptr.1829
Ghaffari H, Ghassam B, Chandra Nayaka S, Ramachandra Kini K, Prakash HS (2014) Antioxidant and neuroprotective activities of Hyptis suaveolens (L.) Poit. against oxidative stress-induced neurotoxicity. Cell Mol Neurobiol 34:323–331. doi:10.1007/s10571-013-0016-7
Guizani N, Waly MI, Ali A, Al-Saidi G, Singh V, Bhatt N, Rahman MS (2011) Papaya epicarp extract protects against hydrogen peroxide-induced oxidative stress in human SH-SY5Y neuronal cells. Exp Biol Med (Maywood) 236:1205–1210. doi:10.1258/ebm.2011.011031
Han BH, Xu D, Choi J, Han Y, Xanthoudakis S, Roy S, Tam J, Vaillancourt J, Colucci J, Siman R, Giroux A, Robertson GS, Zamboni R, Nicholson DW, Holtzman DM (2002) Selective, reversible caspase-3 inhibitor is neuroprotective and reveals distinct pathways of cell death after neonatal hypoxic-ischemic brain injury. J Biol Chem 277:30128–30136. doi:10.1074/jbc.M202931200
Hyam SR et al (2013) Arctigenin ameliorates inflammation in vitro and in vivo by inhibiting the PI3 K/AKT pathway and polarizing M1 macrophages to M2-like macrophages. Eur J Pharmacol 708:21–29. doi:10.1016/j.ejphar.2013.01.014
Jomova K, Valko M (2011) Importance of iron chelation in free radical-induced oxidative stress and human disease. Curr Pharm Des 17:3460–3473. doi:10.2174/138161211798072463
Ju H-Y, Chen SC, Wu K-J, Kuo H-C, Hseu Y-C, Ching H, Wu C-R (2012) Antioxidant phenolic profile from ethyl acetate fraction of Fructus Ligustri Lucidi with protection against hydrogen peroxide-induced oxidative damage in SH-SY5Y cells. Food Chem Toxicol 50:492–502. doi:10.1016/j.fct.2011.11.036
Kim S, Park SE, Sapkota K, Kim MK, Kim SJ (2011) Leaf extract of Rhus verniciflua Stokes protects dopaminergic neuronal cells in a rotenone model of Parkinson’s disease. J Pharm Pharmacol 63:1358–1367. doi:10.1111/j.2042-7158.2011.01342.x
Kumar KH, Khanum F (2013) Hydroalcoholic extract of Cyperus rotundus ameliorates H2O2-induced human neuronal cell damage via its anti-oxidative and anti-apoptotic machinery. Cell Mol Neurobiol 33:5–17. doi:10.1007/s10571-012-9865-8
Kwon S-H, Kim J-A, Hong S-I, Jung Y-H, Kim H-C, Lee S-Y, Jang C-G (2011) Loganin protects against hydrogen peroxide-induced apoptosis by inhibiting phosphorylation of JNK, p38, and ERK 1/2 MAPKs in SH-SY5Y cells. Neurochem Int 58:533–541. doi:10.1016/j.neuint.2011.01.012
Le DA, Wu Y, Huang Z, Matsushita K, Plesnila N, Augustinack JC, Hyman BT, Yuan J, Kuida K, Flavell RA, Moskowitz MA (2002) Caspase activation and neuroprotection in caspase-3- deficient mice after in vivo cerebral ischemia and in vitro oxygen glucose deprivation. Proc Natl Acad Sci U S A 99:15188–15193. doi:10.1073/pnas.232473399
Luo T, Zhang H, Zhang WW, Huang JT, Song EL, Chen SG, He F, Xu J, Wang HQ (2011) Neuroprotective effect of jatrorrhizine on hydrogen peroxide-induced cell injury and its potential mechanisms in PC12 cells. Neurosci Lett 498:227–231. doi:10.1016/j.neulet.2011.05.017
Menazza S, Blaauw B, Tiepolo T, Toniolo L, Braghetta P, Spolaore B, Reggiani C, Di Lisa F, Bonaldo P, Canton M (2010) Oxidative stress by monoamine oxidases is causally involved in myofiber damage in muscular dystrophy. Hum Mol Genet 19:4207–4215. doi:10.1093/hmg/ddq339
Nelson SK, Bose S, Rizeq M, McCord JM (2005) Oxidative stress in organ preservation: a multifaceted approach to cardioplegia. Biomed Pharmacother 59:149–157. doi:10.1016/j.biopha.2005.03.007
Park SY, Hong SS, Han XH, Hwang JS, Lee D, Ro JS, Hwang BY (2007) Lignans from Arctium lappa and their inhibition of LPS-induced nitric oxide production. Chem Pharm Bull (Tokyo) 55:150–152. doi:10.1248/cpb.55.150
Park SE, Kim S, Sapkota K, Kim SJ (2010) Neuroprotective effect of Rosmarinus officinalis extract on human dopaminergic cell line, SH-SY5Y. Cell Mol Neurobiol 30:759–767. doi:10.1007/s10571-010-9502-3
Qian J, Jiang F, Wang B, Yu Y, Zhang X, Yin Z, Liu C (2010) Ophiopogonin D prevents H2O2-induced injury in primary human umbilical vein endothelial cells. J Ethnopharmacol 128:438–445. doi:10.1016/j.jep.2010.01.031
Si CL, Shen T, Jiang YY, Wu L, Yu GJ, Ren XD, Xu GH, Hu WC (2013) Antioxidant properties and neuroprotective effects of isocampneoside II on hydrogen peroxide-induced oxidative injury in PC12 cells. Food Chem Toxicol 59:145–152. doi:10.1016/j.fct.2013.05.051
Sohn EH, Jang SA, Joo H, Park S, Kang SC, Lee CH, Kim SY (2011) Anti-allergic and anti-inflammatory effects of butanol extract from Arctium Lappa L. Clin Mol Allergy 9:4. doi:10.1186/1476-7961-9-4
Somayajulu M, McCarthy S, Hung M, Sikorska M, Borowy-Borowski H, Pandey S (2005) Role of mitochondria in neuronal cell death induced by oxidative stress; neuroprotection by coenzyme Q10. Neurobiol Dis 18:618–627. doi:10.1016/j.nbd.2004.10.021
Tan JW, Tham CL, Israf DA, Lee SH, Kim MK (2013) Neuroprotective effects of biochanin A against glutamate-induced cytotoxicity in PC12 cells via apoptosis inhibition. Neurochem Res 38:512–518. doi:10.1007/s11064-012-0943-6
Tian X, Sui S, Huang J, Bai J-P, Ren T-S, Zhao Q-C (2014) Neuroprotective effects of Arctium lappa L. roots against glutamate-induced oxidative stress by inhibiting phosphorylation of p38, JNK and ERK 1/2 MAPKs in PC12 cells. Environ Toxicol Phar 38:189–198. doi:10.1016/j.etap.2014.05.017
Valko M, Leibfritz D, Moncol J, Cronin MTD, Mazur M, Telser J (2007) Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell B 39:44–84. doi:10.1016/j.biocel.2006.07.001
Venuprasad MP, Hemanth Kumar K, Khanum F (2013) Neuroprotective effects of hydroalcoholic extract of Ocimum sanctum against H2O2 induced neuronal cell damage in SH-SY5Y cells via its antioxidative defence mechanism. Neurochem Res 38:2190–2200. doi:10.1007/s11064-013-1128-7
Wang K, Zhu L, Zhu X, Zhang K, Huang B, Zhang J, Zhang Y, Zhu L, Zhou B, Zhou F (2014) Protective effect of paeoniflorin on Aβ25–35-induced SH-SY5Y cell injury by preventing mitochondrial dysfunction. Cell Mol Neurobiol 34:227–234. doi:10.1007/s10571-013-0006-9
Yan MH, Wang X, Zhu X (2013) Mitochondrial defects and oxidative stress in Alzheimer disease and Parkinson disease. Free Radic Biol Med 62:90–101. doi:10.1016/j.freeradbiomed.2012.11.014
Acknowledgments
This project was supported by the Key National Science & Technology Specific Project of China (2014ZX09J14101-05C).
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**ng Tian, Li-** Guo, **ao-Long Hu, ** Huang, Yan-Hua Fan, Tian-Shu Ren and Qing-Chun Zhao declare that there is no conflict of interest.
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Tian, X., Guo, LP., Hu, XL. et al. Protective Effects of Arctium lappa L. Roots Against Hydrogen Peroxide-Induced Cell Injury and Potential Mechanisms in SH-SY5Y Cells. Cell Mol Neurobiol 35, 335–344 (2015). https://doi.org/10.1007/s10571-014-0129-7
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DOI: https://doi.org/10.1007/s10571-014-0129-7