n-Butylidenephthalide Protects against Dopaminergic Neuron Degeneration and α-Synuclein Accumulation in Caenorhabditis elegans Models of Parkinson's Disease
Background Parkinson's disease (PD) is the second most common degenerative disorder of the central nervous system that impairs motor skills and cognitive function. To date, the disease has no effective therapies. The identification of new drugs that provide benefit in arresting the decline seen in PD patients is the focus of much recent study. However, the lengthy time frame for the progression of neurodegeneration in PD increases both the time and cost of examining potential therapeutic compounds in mammalian models. An alternative is to first evaluate the efficacy of compounds in Caenorhabditis elegans models, which reduces examination time from months to days. n-Butylidenephthalide is the naturally-occurring component derived from the chloroform extract of Angelica sinensis. It has been shown to have anti-tumor and anti-inflammatory properties, but no reports have yet described the effects of n-butylidenephthalide on PD. The aim of this study was to assess the potential for n-butylidenephthalide to improve PD in C. elegans models. Methodology/Principal Findings In the current study, we employed a pharmacological strain that expresses green fluorescent protein specifically in dopaminergic neurons (BZ555) and a transgenic strain that expresses human α-synuclein in muscle cells (OW13) to investigate the antiparkinsonian activities of n-butylidenephthalide. Our results demonstrate that in PD animal models, n-butylidenephthalide significantly attenuates dopaminergic neuron degeneration induced by 6-hydroxydopamine; reduces α-synuclein accumulation; recovers lipid content, food-sensing behavior, and dopamine levels; and prolongs life-span of 6-hydroxydopamine treatment, thus revealing its potential as a possible antiparkinsonian drug. n-Butylidenephthalide may exert its effects by blocking egl-1 expression to inhibit apoptosis pathways and by raising rpn-6 expression to enhance the activity of proteasomes. Conclusions/Significance n-Butylidenephthalide may be one of the effective neuroprotective agents for PD.
Auluck PK, Chan HY, Trojanowski JQ, Lee VM, Bonini NM (2002) Chaperone suppression of alpha-synuclein toxicity in a Drosophila model for Parkinson's disease. Science 295: 865–868.
[6]
Snyder H, Mensah K, Theisler C, Lee J, Matouschek A, et al. (2003) Aggregated and monomeric alpha-synuclein bind to the S6' proteasomal protein and inhibit proteasomal function. J Biol Chem 278: 11753–11759.
[7]
Cuervo AM, Stefanis L, Fredenburg R, Lansbury PT, Sulzer D (2004) Impaired degradation of mutant alpha-synuclein by chaperone-mediated autophagy. Science 305: 1292–1295.
[8]
Bussell R Jr, Eliezer D (2004) Effects of Parkinson's disease-linked mutations on the structure of lipid-associated alpha-synuclein. Biochemistry 43: 4810–4818.
[9]
Jensen PH, Nielsen MS, Jakes R, Dotti CG, Goedert M (1998) Binding of alpha-synuclein to brain vesicles is abolished by familial Parkinson's disease mutation. J Biol Chem 273: 26292–26294.
[10]
Binukumar BK, Bal A, Kandimalla RJ, Gill KD (2010) Nigrostriatal neuronal death following chronic dichlorvos exposure: crosstalk between mitochondrial impairments, alpha synuclein aggregation, oxidative damage and behavioral changes. Mol Brain 3: 35.
[11]
Jankovic J, Poewe W (2012) Therapies in Parkinson's disease. Curr Opin Neurol 25: 433–447.
[12]
Yi L, Liang Y, Wu H, Yuan D (2009) The analysis of Radix Angelicae Sinensis (Danggui). J Chromatogr A 1216: 1991–2001.
[13]
Chiu SC, Chen SP, Huang SY, Wang MJ, Lin SZ, et al. (2012) Induction of apoptosis coupled to endoplasmic reticulum stress in human prostate cancer cells by n-butylidenephthalide. PLoS One 7: e33742.
[14]
Wei CW, Lin CC, Yu YL, Lin CY, Lin PC, et al. (2009) n-Butylidenephthalide induced apoptosis in the A549 human lung adenocarcinoma cell line by coupled down-regulation of AP-2alpha and telomerase activity. Acta Pharmacol Sin 30: 1297–1306.
[15]
Chen YL, Jian MH, Lin CC, Kang JC, Chen SP, et al. (2008) The induction of orphan nuclear receptor Nur77 expression by n-butylenephthalide as pharmaceuticals on hepatocellular carcinoma cell therapy. Mol Pharmacol 74: 1046–1058.
[16]
Yeh JC, Cindrova-Davies T, Belleri M, Morbidelli L, Miller N, et al. (2011) The natural compound n-butylidenephthalide derived from the volatile oil of Radix Angelica sinensis inhibits angiogenesis in vitro and in vivo. Angiogenesis 14: 187–197.
[17]
Fu RH, Hran HJ, Chu CL, Huang CM, Liu SP, et al. (2011) Lipopolysaccharide-stimulated activation of murine DC2.4 cells is attenuated by n-butylidenephthalide through suppression of the NF-kappaB pathway. Biotechnol Lett 33: 903–910.
[18]
Teng CM, Chen WY, Ko WC, Ouyang CH (1987) Antiplatelet effect of butylidenephthalide. Biochim Biophys Acta 924: 375–382.
[19]
Ko WC, Liao CC, Shih CH, Lei CB, Chen CM (2002) Relaxant effects of butylidenephthalide in isolated dog blood vessels. Planta Med 68: 1004–1009.
[20]
Chan SS, Choi AO, Jones RL, Lin G (2006) Mechanisms underlying the vasorelaxing effects of butylidenephthalide, an active constituent of Ligusticum chuanxiong, in rat isolated aorta. Eur J Pharmacol 537: 111–117.
[21]
Chan SS, Jones RL, Lin G (2009) Synergistic interaction between the Ligusticum chuanxiong constituent butylidenephthalide and the nitric oxide donor sodium nitroprusside in relaxing rat isolated aorta. J Ethnopharmacol 122: 308–312.
[22]
Ko WC, Sheu JR, Tzeng SH, Chen CM (1998) The selective antianginal effect without changing blood pressure of butylidenephthalide in conscious rats. Planta Med 64: 229–232.
[23]
Ko WC, Charng CY, Sheu JR, Tzeng SH, Chen CM (1998) Effect of butylidenephthalide on calcium mobilization in isolated rat aorta. J Pharm Pharmacol 50: 1365–1369.
[24]
Mimura Y, Kobayashi S, Naitoh T, Kimura I, Kimura M (1995) The structure-activity relationship between synthetic butylidenephthalide derivatives regarding the competence and progression of inhibition in primary cultures proliferation of mouse aorta smooth muscle cells. Biol Pharm Bull 18: 1203–1206.
[25]
Harrington AJ, Yacoubian TA, Slone SR, Caldwell KA, Caldwell GA (2012) Functional analysis of VPS41-mediated neuroprotection in Caenorhabditis elegans and mammalian models of Parkinson's disease. J Neurosci 32: 2142–2153.
[26]
Kuwahara T, Tonegawa R, Ito G, Mitani S, Iwatsubo T (2012) Phosphorylation of alpha-synuclein protein at Ser-129 reduces neuronal dysfunction by lowering its membrane binding property in Caenorhabditis elegans. J Biol Chem 287: 7098–7109.
[27]
Jadiya P, Chatterjee M, Sammi SR, Kaur S, Palit G, et al. (2011) Sir-2.1 modulates 'calorie-restriction-mediated' prevention of neurodegeneration in Caenorhabditis elegans: implications for Parkinson's disease. Biochem Biophys Res Commun 413: 306–310.
[28]
Liu Z, Hamamichi S, Lee BD, Yang D, Ray A, et al. (2011) Inhibitors of LRRK2 kinase attenuate neurodegeneration and Parkinson-like phenotypes in Caenorhabditis elegans and Drosophila Parkinson's disease models. Hum Mol Genet 20: 3933–3942.
[29]
Schreiber MA, McIntire SL (2011) A Caenorhabditis elegans p38 MAP kinase pathway mutant protects from dopamine, methamphetamine, and MDMA toxicity. Neurosci Lett 498: 99–103.
[30]
Martinez-Finley EJ, Avila DS, Chakraborty S, Aschner M (2011) Insights from Caenorhabditis elegans on the role of metals in neurodegenerative diseases. Metallomics 3: 271–279.
[31]
Benedetto A, Au C, Avila DS, Milatovic D, Aschner M (2010) Extracellular dopamine potentiates mn-induced oxidative stress, lifespan reduction, and dopaminergic neurodegeneration in a BLI-3-dependent manner in Caenorhabditis elegans. PLoS Genet 6: e1001084.
[32]
Yao C, El Khoury R, Wang W, Byrd TA, Pehek EA, et al. (2010) LRRK2-mediated neurodegeneration and dysfunction of dopaminergic neurons in a Caenorhabditis elegans model of Parkinson's disease. Neurobiol Dis 40: 73–81.
[33]
Kamp F, Exner N, Lutz AK, Wender N, Hegermann J, et al. (2010) Inhibition of mitochondrial fusion by alpha-synuclein is rescued by PINK1, Parkin and DJ-1. EMBO J 29: 3571–3589.
Asikainen S, Rudgalvyte M, Heikkinen L, Louhiranta K, Lakso M, et al. (2010) Global microRNA expression profiling of Caenorhabditis elegans Parkinson's disease models. J Mol Neurosci 41: 210–218.
[36]
Roodveldt C, Bertoncini CW, Andersson A, van der Goot AT, Hsu ST, et al. (2009) Chaperone proteostasis in Parkinson's disease: stabilization of the Hsp70/alpha-synuclein complex by Hip. EMBO J 28: 3758–3770.
[37]
Ruan Q, Harrington AJ, Caldwell KA, Caldwell GA, Standaert DG (2010) VPS41, a protein involved in lysosomal trafficking, is protective in Caenorhabditis elegans and mammalian cellular models of Parkinson's disease. Neurobiol Dis 37: 330–338.
[38]
Caldwell KA, Tucci ML, Armagost J, Hodges TW, Chen J, et al. (2009) Investigating bacterial sources of toxicity as an environmental contributor to dopaminergic neurodegeneration. PLoS One 4: e7227.
[39]
Bargmann CI (1998) Neurobiology of the Caenorhabditis elegans genome. Science 282: 2028–2033.
[40]
van der Goot AT, Zhu W, Vazquez-Manrique RP, Seinstra RI, Dettmer K, et al. (2012) Delaying aging and the aging-associated decline in protein homeostasis by inhibition of tryptophan degradation. Proc Natl Acad Sci U S A 109: 14912–14917.
[41]
Lee JY, Song J, Kwon K, Jang S, Kim C, et al. (2012) Human DJ-1 and its homologs are novel glyoxalases. Hum Mol Genet 21: 3215–3225.
[42]
Zheng M, Gorelenkova O, Yang J, Feng Z (2012) A liquid phase based C. elegans behavioral analysis system identifies motor activity loss in a nematode Parkinson's disease model. J Neurosci Methods 204: 234–237.
[43]
Vidal-Gadea AG, Pierce-Shimomura JT (2012) Conserved role of dopamine in the modulation of behavior. Commun Integr Biol 5: 440–447.
[44]
Kuwahara T, Koyama A, Koyama S, Yoshina S, Ren CH, et al. (2008) A systematic RNAi screen reveals involvement of endocytic pathway in neuronal dysfunction in alpha-synuclein transgenic C. elegans. Hum Mol Genet 17: 2997–3009.
De Jesus-Cortes H, Xu P, Drawbridge J, Estill SJ, Huntington P, et al. (2012) Neuroprotective efficacy of aminopropyl carbazoles in a mouse model of Parkinson disease. Proc Natl Acad Sci U S A 109: 17010–17015.
[47]
Cao P, Yuan Y, Pehek EA, Moise AR, Huang Y, et al. (2010) Alpha-synuclein disrupted dopamine homeostasis leads to dopaminergic neuron degeneration in Caenorhabditis elegans. PLoS One 5: e9312.
[48]
Slone SR, Lesort M, Yacoubian TA (2011) 14-3-3theta protects against neurotoxicity in a cellular Parkinson's disease model through inhibition of the apoptotic factor Bax. PLoS One 6: e21720.
[49]
Harrington AJ, Knight AL, Caldwell GA, Caldwell KA (2011) Caenorhabditis elegans as a model system for identifying effectors of alpha-synuclein misfolding and dopaminergic cell death associated with Parkinson's disease. Methods 53: 220–225.
[50]
Yacoubian TA, Slone SR, Harrington AJ, Hamamichi S, Schieltz JM, et al. (2010) Differential neuroprotective effects of 14-3-3 proteins in models of Parkinson's disease. Cell Death Dis 1: e2.
[51]
Jadiya P, Khan A, Sammi SR, Kaur S, Mir SS, et al. (2011) Anti-Parkinsonian effects of Bacopa monnieri: insights from transgenic and pharmacological Caenorhabditis elegans models of Parkinson's disease. Biochem Biophys Res Commun 413: 605–610.
[52]
Locke CJ, Fox SA, Caldwell GA, Caldwell KA (2008) Acetaminophen attenuates dopamine neuron degeneration in animal models of Parkinson's disease. Neurosci Lett 439: 129–133.
[53]
Fu RH, Wang YC, Chen CS, Tsai RT, Liu SP, et al.. (2013) Acetylcorynoline attenuates dopaminergic neuron degeneration and alpha-synuclein aggregation in animal models of Parkinson's disease. Neuropharmacology doi:10.1016/j.neuropharm.2013.08.007.
[54]
Brenner S (1974) The genetics of Caenorhabditis elegans. Genetics 77: 71–94.
[55]
Yang X, Zhang P, Wu J, Xiong S, Jin N, et al. (2012) The neuroprotective and lifespan-extension activities of Damnacanthus officinarum extracts in Caenorhabditis elegans. J Ethnopharmacol 141: 41–47.
[56]
Voisine C, Varma H, Walker N, Bates EA, Stockwell BR, et al. (2007) Identification of potential therapeutic drugs for huntington's disease using Caenorhabditis elegans. PLoS One 2: e504.
[57]
Nass R, Hall DH, Miller DM 3rd, Blakely RD (2002) Neurotoxin-induced degeneration of dopamine neurons in Caenorhabditis elegans. Proc Natl Acad Sci U S A 99: 3264–3269.
[58]
Sawin ER, Ranganathan R, Horvitz HR (2000) C. elegans locomotory rate is modulated by the environment through a dopaminergic pathway and by experience through a serotonergic pathway. Neuron 26: 619–631.
[59]
Kuwahara T, Koyama A, Gengyo-Ando K, Masuda M, Kowa H, et al. (2006) Familial Parkinson mutant alpha-synuclein causes dopamine neuron dysfunction in transgenic Caenorhabditis elegans. J Biol Chem 281: 334–340.
[60]
Hoogewijs D, Houthoofd K, Matthijssens F, Vandesompele J, Vanfleteren JR (2008) Selection and validation of a set of reliable reference genes for quantitative sod gene expression analysis in C. elegans. BMC Mol Biol 9: 9.
[61]
Vilchez D, Morantte I, Liu Z, Douglas PM, Merkwirth C, et al. (2012) RPN-6 determines C. elegans longevity under proteotoxic stress conditions. Nature 489: 263–268.
[62]
Shvadchak VV, Yushchenko DA, Pievo R, Jovin TM (2011) The mode of alpha-synuclein binding to membranes depends on lipid composition and lipid to protein ratio. FEBS Lett 585: 3513–3519.
[63]
Bove J, Perier C (2012) Neurotoxin-based models of Parkinson's disease. Neuroscience 211: 51–76.
[64]
Saw CL, Wu Q, Su ZY, Wang H, Yang Y, et al.. (2013) Effects of natural phytochemicals in Angelica sinensis (Danggui) on Nrf2-mediated gene expression of phase II drug metabolizing enzymes and anti-inflammation. Biopharm Drug Dispos DOI: 10.1002/bdd.1846.
[65]
Hwang CK, Chun HS (2012) Isoliquiritigenin isolated from licorice Glycyrrhiza uralensis prevents 6-hydroxydopamine-induced apoptosis in dopaminergic neurons. Biosci Biotechnol Biochem 76: 536–543.
[66]
Charriaut-Marlangue C (2004) Apoptosis: a target for neuroprotection. Therapie 59: 185–190.
[67]
Pourkarimi E, Greiss S, Gartner A (2012) Evidence that CED-9/Bcl2 and CED-4/Apaf-1 localization is not consistent with the current model for C. elegans apoptosis induction. Cell Death Differ 19: 406–415.
[68]
Hyman BT, Yuan J (2012) Apoptotic and non-apoptotic roles of caspases in neuronal physiology and pathophysiology. Nat Rev Neurosci 13: 395–406.
[69]
van Ham TJ, Thijssen KL, Breitling R, Hofstra RM, Plasterk RH, et al. (2008) C. elegans model identifies genetic modifiers of alpha-synuclein inclusion formation during aging. PLoS Genet 4: e1000027.
[70]
Hamamichi S, Rivas RN, Knight AL, Cao S, Caldwell KA, et al. (2008) Hypothesis-based RNAi screening identifies neuroprotective genes in a Parkinson's disease model. Proc Natl Acad Sci U S A 105: 728–733.
[71]
Karpinar DP, Balija MB, Kugler S, Opazo F, Rezaei-Ghaleh N, et al. (2009) Pre-fibrillar alpha-synuclein variants with impaired beta-structure increase neurotoxicity in Parkinson's disease models. EMBO J 28: 3256–3268.
[72]
Usenovic M, Knight AL, Ray A, Wong V, Brown KR, et al. (2012) Identification of novel ATP13A2 interactors and their role in alpha-synuclein misfolding and toxicity. Hum Mol Genet 21: 3785–3794.
[73]
van Ham TJ, Holmberg MA, van der Goot AT, Teuling E, Garcia-Arencibia M, et al. (2010) Identification of MOAG-4/SERF as a regulator of age-related proteotoxicity. Cell 142: 601–612.
[74]
Nedelsky NB, Todd PK, Taylor JP (2008) Autophagy and the ubiquitin-proteasome system: collaborators in neuroprotection. Biochim Biophys Acta 1782: 691–699.
[75]
Yu Z, Kleifeld O, Lande-Atir A, Bsoul M, Kleiman M, et al. (2011) Dual function of Rpn5 in two PCI complexes, the 26S proteasome and COP9 signalosome. Mol Biol Cell 22: 911–920.
[76]
Maccarrone M, Bernardi G, Agro AF, Centonze D (2011) Cannabinoid receptor signalling in neurodegenerative diseases: a potential role for membrane fluidity disturbance. Br J Pharmacol 163: 1379–1390.
[77]
Sebastiao AM, Colino-Oliveira M, Assaife-Lopes N, Dias RB, Ribeiro JA (2013) Lipid rafts, synaptic transmission and plasticity: impact in age-related neurodegenerative diseases. Neuropharmacology 64: 97–107.
[78]
Hirsch EC, Hunot S (2009) Neuroinflammation in Parkinson's disease: a target for neuroprotection? Lancet Neurol 8: 382–397.
[79]
Rogers J, Mastroeni D, Leonard B, Joyce J, Grover A (2007) Neuroinflammation in Alzheimer's disease and Parkinson's disease: are microglia pathogenic in either disorder? Int Rev Neurobiol 82: 235–246.
[80]
Collins LM, Toulouse A, Connor TJ, Nolan YM (2012) Contributions of central and systemic inflammation to the pathophysiology of Parkinson's disease. Neuropharmacology 62: 2154–2168.
[81]
Kim HG, Ju MS, Ha SK, Lee H, Kim SY, et al. (2012) Acacetin protects dopaminergic cells against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyri？dine-inducedneuroinflammation in vitro and in vivo. Biol Pharm Bull 35: 1287–1294.
[82]
Du XX, Xu HM, Jiang H, Song N, Wang J, et al. (2012) Curcumin protects nigral dopaminergic neurons by iron-chelation in the 6-hydroxydopamine rat model of Parkinson's disease. Neurosci Bull 28: 253–258.
[83]
Kumar H, Kim IS, More SV, Kim BW, Bahk YY, et al. (2013) Gastrodin protects apoptotic dopaminergic neurons in a toxin-induced Parkinson's disease model. Evid Based Complement Alternat Med 2013: 514095.
[84]
Zhang ZJ, Cheang LC, Wang MW, Lee SM (2011) Quercetin exerts a neuroprotective effect through inhibition of the iNOS/NO system and pro-inflammation gene expression in PC12 cells and in zebrafish. Int J Mol Med 27: 195–203.
[85]
Lu JH, Tan JQ, Durairajan SS, Liu LF, Zhang ZH, et al. (2012) Isorhynchophylline, a natural alkaloid, promotes the degradation of alpha-synuclein in neuronal cells via inducing autophagy. Autophagy 8: 98–108.
[86]
Zbarsky V, Datla KP, Parkar S, Rai DK, Aruoma OI, et al. (2005) Neuroprotective properties of the natural phenolic antioxidants curcumin and naringenin but not quercetin and fisetin in a 6-OHDA model of Parkinson's disease. Free Radic Res 39: 1119–1125.
[87]
Ramassamy C (2006) Emerging role of polyphenolic compounds in the treatment of neurodegenerative diseases: a review of their intracellular targets. Eur J Pharmacol 545: 51–64.
[88]
Pandey N, Strider J, Nolan WC, Yan SX, Galvin JE (2008) Curcumin inhibits aggregation of alpha-synuclein. Acta Neuropathol 115: 479–489.
[89]
Ahmad B, Lapidus LJ (2012) Curcumin prevents aggregation in alpha-synuclein by increasing reconfiguration rate. J Biol Chem 287: 9193–9199.
[90]
Berman AE, Chan WY, Brennan AM, Reyes RC, Adler BL, et al. (2011) N-acetylcysteine prevents loss of dopaminergic neurons in the EAAC1-/- mouse. Ann Neurol 69: 509–520.
[91]
Clark J, Clore EL, Zheng K, Adame A, Masliah E, et al. (2010) Oral N-acetyl-cysteine attenuates loss of dopaminergic terminals in alpha-synuclein overexpressing mice. PLoS One 5: e12333.
[92]
Etminan M, Gill SS, Samii A (2005) Intake of vitamin E, vitamin C, and carotenoids and the risk of Parkinson's disease: a meta-analysis. Lancet Neurol 4: 362–365.
[93]
Tsai NM, Chen YL, Lee CC, Lin PC, Cheng YL, et al. (2006) The natural compound n-butylidenephthalide derived from Angelica sinensis inhibits malignant brain tumor growth in vitro and in vivo. J Neurochem 99: 1251–1262.
[94]
Tan W, Li Y, Chen M, Wang Y (2011) Berberine hydrochloride: anticancer activity and nanoparticulate delivery system. Int J Nanomedicine 6: 1773–1777.
[95]
Onishi H, Machida Y (2005) Macromolecular and nanotechnological modification of camptothecin and its analogs to improve the efficacy. Curr Drug Discov Technol 2: 169–183.
[96]
Fu RH, Liu SP, Ou CW, Huang CM, Wang YC (2010) Spatial control of cells, peptide delivery and dynamic monitoring of cellular physiology with chitosan-assisted dual color quantum dot FRET peptides. Acta Biomater 6: 3621–3629.
