Therapeutic Potential of Citrus sinensis Peels Against Rotenone Induced Parkinsonism in Rats

Author(s): Manal Hamed*, Asmaa Aboul Naser, Marwa Elbatanony, Amal El-Feky, Azza Matloub, Nagy El-Rigal, Wagdy Khalil

Journal Name: Current Bioactive Compounds

Volume 17 , Issue 6 , 2021

Article ID: e010621186105
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Graphical Abstract:


Background: Parkinson’s disease (PD) is one of the most common neurodegenerative disorders spread worldwide in elderly people.

Methods: The Citrus peels methanolic extract (100 mg/kg body weight) was evaluated as an antiparkinsonism agent in rats through estimation of oxidative stress markers, neurotransmitter levels, energetic indices, DNA fragmentation pattern, inflammatory mediators, adenosine A2A receptor gene expression and the histopathological analysis of the brain. In addition, its effect was compared with ZM241385; an adenosine A2A receptor antagonist, as well as the classical drug; (L-dopa).

Results: The methanolic extract of C. sinensis peels constituted 17.59 ± 1.92 mg GAE/g and 4.88 ± 0.43 mg CE/g of total phenolic and flavonoid content, respectively. The polyphenolic composition was qualified and quantified using HPLC/DAD and UPLC/ESI-MS analysis. HPLC/DAD analysis led to identify 8 phenolic acids and 4 flavonoids. UPLC/MS analysis led to identify 20 polyphenolic compounds, including 9 polymethoxylated flavoniods, 7 flavonoidal glycosides and 4 phenolic derivatives. Nobiletin and tangeretin were found as abundant polymethoxylated flavones while, hesperidin and 1-caffeoyl-β-D-glucose were found as abundant glycosyl flavone and phenolic derivatives, respectively. Rotenone induced rats showed a significant decrease in neurotransmitter levels, energetic and antioxidant parameters, while a significant increase in total protein, inflammatory mediators, adenosine A2A receptor gene expression, DNA and lipid peroxidation levels was recorded. Treatments with plant extract, L-dopa and ZM241385 restored these selected parameters to variable extents with a more potent effect of ZM241385 than L-dopa. Rotenone induced rats were left free without treatment; not recorded a noticeable improvement level.

Conclusion: Citrus sinensis peels was rich with bioactive valuable-added products. This may lead to the development of new nutraceutical and pharmaceutical agents as well as functional food products used as anti-oxidative, anti-inflammatory and anti-parkinsonian agent.

Keywords: Citrus sinensis peels, polymethoxylated flavones, hesperidin, parkinson's disease, ZM241385, L-dopa.

Perez CA, Tong Y, Guo M. Iron chelators as potential therapeutic agents for Parkinson’s disease. Curr Bioact Compd 2008; 4: 150-8.
Lebouvier T. The second brain and Parkinson’s disease. Eur J Neurosci 2009; 30: 735-41.
Reyhani-Rad S, Nayebi AM, Mahmoudi J, Samini M. Role of 5-Hydroxytryptamine 1A receptors in 6-hydroxydopmaine-induced catalepsy-like immobilization in rats: a therapeutic approach for treating catalepsy of Parkinson’s disease. Iran J Pharm Res 2012; 11: 1175-81.
Mahmoudi J, Nayebi AM, Reyhani-Rad S, Samini M. Fluoxetine improves the effect of levodopa on 6-hydroxy dopamine-induced motor impairments in rats. Adv Pharm Bull 2012; 2: 149-55.
Hunot S, Hirsch EC. Neuroinflammatory processes in Parkinson’s disease. Ann Neurol 2003; 53: S49-60.
Abbott NJ, Ronnback L, Hansson E. Astrocyte-endothelial interactions at the blood-brain barrier. Nat Rev Neurosci 2006; 7: 41-53.
Sherer TB, Betarbet R, Testa CM, Seo BB, Richardso JR, Kim JH. Mechanism of toxicity in rotenone models of Parkinson’s disease. J Neurosci 2003; 34: 10756-64.
Gao HM, Liu B, Hong JS. Critical role for microglial NADPH oxidase in rotenone-induced degeneration of dopaminergic neurons. J Neurosci 2003; 23: 6181-7.
Freestone PS, Chung KK, Guatteo E, Mercuri NB, Nicholson LF, Lipski J. Acute action of rotenone on nigral dopaminergic neurons-involvement of reactive oxygen species and disruption of Ca2+homeostasis. Eur J Neurosci 2009; 30: 1849-59.
Yong R, Liu RW, Jiang H, Jiang Q, Feng J. Selective vulnerability of dopaminergic neurons to microtubule depolymerization. J Biol Chem 2005; 280: 34105-12.
Priyanga SK. VijayaIakshmi, K.; Selvaraj, R. Behavioural studies of Wistar rates in rotenone induce model of Parkinson’s disease. Int J Pharm Pharm Sci 2017; 9: 159-64.
Dorszewska J, Prendecki M, Lianeri M, Kozubski W. Molecular effects of L-dopa therapy in Parkinson’s disease. Curr Genet 2014; 15: 11-7.
Fathalla AM, Soliman AM, Ali MH, Moustafa AA. Adenosine A2A receptor blockade prevents rotenone-induced motor impairment in a rat model of Parkinsonism. Front Behav Neurosci 2016; 10: 1-5.
Khan SA, Ahmad R, Asad SA, Muhammad S. Citrus Flavonoids: Their Biosynthesis, Functions and Genetic Improvement.Molecular Characterization of Citrus Cultivars: Insight from Recent Studies Bernardi J Adriano Marocco A Caruso P Licciardello C New York Nova Science Publishers. 2014.
Anagnostopoulou MA, Kefalas P, Kokkalou E, Assimopoulou AN, Papageorgiou VP. Analysis of antioxidant compounds in sweet orange peel by HPLC-diode array detection-electrospray ionization mass spectrometry. Biomed Chromatogr 2005; 19: 138-48.
Li S. Lo; C.-Y.; Ho, C.-T. Hydroxylated polymethoxyflavones and methylated flavonoids in sweet orange (Citrus sinensis) peel. J Agric Food Chem 2006; 54: 4176-85.
Weber B, Hartmann B, Stöckigt D, et al. Liquid chromatography/mass spectrometry and liquid chromatography/nuclear magnetic resonance as complementary analytical techniques for unambiguous identification of polymethoxylated flavones in residues from molecular distillation of orange peel oils (Citrus sinensis). J Agric Food Chem 2006; 54: 274-8.
Mahato N, Sinha M, Sharma K, Koteswararao R, Cho MH. Modern extraction and purification techniques for obtaining high purity food-grade bioactive compounds and value-added co-products from citrus wastes. Foods 2019; 8: 523.
Iglesias-Carres L, Mas-Capdevila A, Bravo FI, Aragonès G, Muguerza B, Arola-Arnal A. Optimization of a polyphenol extraction method for sweet orange pulp (Citrus sinensis L.) to identify phenolic compounds consumed from sweet oranges. PLoS One 2019; 14: e0211267.
Liew SS, Ho WY, Yeap SK, Sharifudin SAB. Phytochemical composition and in vitro antioxidant activities of Citrus sinensis peel extracts. PeerJ 2018; 6: e5331.
Setyawati I, Anggraeni R. Effectiveness of sweet orange peel extract (Citrus sinensis) on the improvement of liver functions of animal trials induced by cigarette smoke. J Young Pharm 2018; 10: s132-5.
Pantsulaia I, Iobadze M, Pantsulaia N, Chikovani T. The effect of citrus peel extracts on cytokines levels and T regulatory cells in acute liver injury. BioMed Res Int 2014; 2014: 127879.
Rafiq S, Kaul R, Sofi SA, Bashir N, Nazir F, Nayik GA. Citrus peel as a source of functional ingredient: a review. J Saudi Soc Agric Sci 2018; 17: 351-8.
Zilic S, Serpen A, Akillioglu G, Jankovic M, Gokmen V. Distributions of phenolic compounds, yellow pigments and oxidative enzymes in wheat grains and their relation to antioxidant capacity of bran and debranned flour. J Cereal Sci 2012; 56: 652-8.
Borai IH, Ezz MK, Rizk MZ, et al. Therapeutic impact of grape leaves polyphenols on certain biochemical and neurological markers in AlCl3-induced Alzheimer’s disease. Biomed Pharmacother 2017; 93: 837-51.
El-Shebiney SA, El-Denshary ES, Abdel-Salam OME, et al. Cannabis resin extract in Parkinson’s disease: Behavioral, neurochemical, and histological evaluation. Cell Biol Res Ther 2014; 3: 1.
Alam M, Schmidt WJ. l-DOPA reverses the hypokinetic behaviour and rigidity in rotenone-treated rats. Behav Brain Res 2004; 153: 439-46.
Sanberg P, Martinez R, Shytle R, Cahill D. The catalepsy test: is a standardized method possible?Motor Activity and Movement Disorders. NY, USA: Humana Press 1996.
Buege JA, Aust SD. Microsomal lipid peroxidation. Methods Enzymol 1978; 52: 302-10.
Moron MS, Depierre JW, Mannervik B. Level of glutathione, glutathione reductase and glutathone-S-transferase activities in rat lung and liver. Biochim Biophys Acta 1972; 582: 67-78.
Nishikimi M, Rae NA, Yagi K. The occurrence of superoxide anion in the action of reduced phenazinemethosulphate and molecular oxygen. Biochem Biophys Res Commun 1972; 46: 849-53.
Wang Y, Yang F, Zhang HX, et al. Cuprous oxide nanoparticles inhibit the growth and metastasis of melanoma by targeting mitochondria Cell Death Dis 2013; 4: e783.
Liu YE, Tong CC, Zhang YB, et al. Chitosan oligosaccharide ameliorates acute lung injury induced by blast injury through the DDAH1/ADMA pathway. PLoS One 2018; 13: e0192135.
Rice ME, Shelton E. Comparison of the reduction of two tetrazolium salts with succinoxidase activity of tissue homogenates. J Natl Cancer Inst 1975; 18: 117-25.
Babson AL, Babson SR. Kinetic colorimetric measurement of serum lactate dehydrogenase activity. Clin Chem 1973; 19: 766-9.
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 1976; 72: 248-54.
Lu T, Xu Y, Mericle MT, Mellgren RL. Participation of the conventional calpains in apoptosis. Biochim Biophys Acta 2002; 1590: 16-26.
El-Baz FK, Khalil WKB, Booles HF, Aly HF, Ali GH. Dunaliella salina suppress oxidative stress, alterations in the expression of pro-apoptosis and inflammation related genes induced by STZ in diabetic rats. Int J Pharm Sci Rev Res 2016; 38: 219-26.
Khalil WKB, Booles HF. Protective role of selenium against over-expression of cancer-related apoptotic genes induced by o-cresol in rats. Arch Indus Hyg Toxicol 2011; 62: 121-9.
Linjawi SAA, Khalil WKB, Salem LM. Detoxified Jatropha curcas kernel meal impact against benzene-induce genetic toxicity in male rats. Int J Pharm 2014; 4: 57-66.
Bancroft J, Stevens A. Theory and Practice of Histological Techniques, Fourthed. Edinburgh, Scotland: Churchil Livingstone 1996.
Hernández-Carranza P, Ávila-Sosa R, Guerrero-Beltrán JA, Navarro-Cruz AR, Corona-Jiménez E, Ochoa-Velasco CE. Optimization of antioxidant compounds extraction from fruit by-products: Apple pomace, orange and banana peel. J Food Process Preserv 2015; 40: 103-15.
Swatsitang P, Tucker G, Robards K, Jardine D. Isolation and identification of phenolic compounds in Citrus sinensis. Anal Chim Acta 2000; 417: 231-40.
Francescato LN, Debenedetti SL, Schwanz TG, Bassani VL, Henriques AT. Identification of phenolic compounds in Equisetum giganteum by LC–ESI-MS/MS and a new approach to total flavonoid quantification. Talanta 2013; 105: 192-203.
Aboul Naser AF, Younis EA, El-Feky AM, Elbatanony MM, Hamed MA. Management of Citrus sinensis peels for protection and treatment against gastric ulcer induced by ethanol in rats. Biomarkers 2020; 25: 349-59.
Cuyckens F, Ma YL, Pocsfalvi G, Claeysi M. Tandem mass spectral strategies for the structural characterization of flavonoid glycosides. Analysis 2000; 28: 888-95.
Kachlicki P, Piasecka A, Stobiecki M, Marczak Ł. Structural characterization of flavonoid glycoconjugates and their derivatives with mass spectrometric techniques. Molecules 2016; 21: 1494.
Sudto K, Pornpakakul S, Wanichwecharungruang S. An efficient method for the large scale isolation of naringin from pomelo (Citrus grandis) peel. Int J Food Sci Technol 2009; 44: 1737-42.
Sharma P, Pandey P, Gupta R. Roshan1, S.; Garg, A.; Shulka, A.; Pasi, A. Isolation and characterization of hesperidin from orange peel. J Pharm Res 2013; 3: 3892-7.
Tang KSC, Konczak I, Zhao J. Identification and quantification of phenolics in Australian native mint (Mentha australis R. Br.). Food Chem 2016; 192: 698-705.
Greenamyre JT, Hastings TG. Parkinson’s: divergentcauses, convergent mechanisms. Science 2004; 304: 1120-2.
Lotharius J, Brundin P. Pathogenesis of Parkinson’s disease: dopamine, vesicles and α-synuclein. Nat Rev Neurosci 2002; 3: 932-42.
Radad K, Rausch WD, Gille G. Rotenone induces cell deathin primary dopaminergic culture by increasing ROS production and inhibiting mitochondrial respiration. Neurochem Int 2006; 49: 379-86.
Keane PC, Kurzawa M, Blain PG, Morris CM. Mitochondrial dysfunction in Parkinson’s disease. Biochim Biophys Acta 2010; 1802: 29-44.
Zaitone S, Abo-Elmatty D, Shaalan A. Acetyl-L-carnitineand α-lipoic acid affect rotenone-induced damage in nigral dopaminergic neurons of rat brain implication for Parkinson’s disease therapy. Pharmacol Biochem Behav 2012; 100: 347-60.
Hamed MA, Aboul Naser AF, Aziz WM, et al. Natural sources, dopaminergic and non-dopaminergic agents for therapeutic assessment of Parkinsonism in rat model. PharmaNutrition 2020; 11 : 100171.
Hamed MA, Mohammed MA, Aboul Naser AF, et al. Optimization of curcuminoids extraction for evaluation against Parkinson’s disease in rats. J Biol Active Prod Nature 2019; 9: 335-51.
Motawi TK, Sadik NAH, Hamed MA, Ali SA, Khalil WKB, Ahmed YR. Potential therapeutic effects of antagonizing adenosine A2A receptor, curcumin and niacin in rotenone-induced Parkinson’s disease mice model. Mol Cell Biochem 2019; 465: 89-102.
Stokes AH, Hastings TG, Vrana KE. Cytotoxic andgenotoxic potential of dopamine. J Neurosci Res 1999; 55: 659-65.
Exner N, Lutz AK, Haass C, Winklhofer KF. Mitochondrial dysfunction in Parkinson’s disease: molecular mechanisms and pathophysiological consequences. Eur Mol Biol Org 2012; 31: 3038-62.
Rothaug M, Becker-Pauly C, Rose-John S. The role of interleukin-6 signaling in nervous tissue. Biochim Biophys Acta 2016; 1863: 1218-27.
Yuan J, Ren J, Wang Y, He X, Zhao Y. Acteoside binds to caspase-3 and exerts neuroprotection in the rotenone rat model of Parkinson’s disease. PLoS One 2016; 11: e0162696.
Ogawa N, Asanuma M, Kondo Y, Kawada Y, Yamamoto M. Differential effects of chronic L-dopa treatment on lipidperoxidation in the mouse brain with or without pretreatmentwith 6-hydroxydopamine. Neurosci Lett 1994; 171: 55-8.
Datla KP, Christidou M, Widmer WW, Rooprai HK, Dexter DT. Tissue distribution and neuroprotective effects of citrus flavonoid tangeretin in a rat model of Parkinson’s disease. Neuroreport 2001; 12: 3871-5.
Jenner P. AnoverviewofadenosineA2A receptor antagonistsin Parkinson’s disease. Int Rev Neurobiol 2014; 119: 71-86.
Pinna A, Bonaventura J, Farré D, Sánchez M, Simola N, Mallol J. L-DOPA disrupts adenosine A2A-cannabinoid CB1 dopamine D2 receptor heteromer cross-talk in the striatum of hemiparkinsonian rats: biochemical and behavioral studies. Exp Neurol 2014; 253: 180-91.
Gyoneva S, Shapiro L, Lazo C, Garnier-Amblard E, Smith Y, Miller GW. Adenosine A2A receptor antagonism reverses inflammation- induced impairment of microglial process extension in a model of Parkinson’s disease. Neurobiol Dis 2014; 67: 191-202.
Hwang S-L, Shih P-H, Yen G-C. Neuroprotective effects of citrus flavonoids. J Agric Food Chem 2012; 60: 877-85.
Sun Y, Han Y, Song M, et al. Inhibitory effects of nobiletin and its major metabolites on lung tumorigenesis. Food Funct 2019; 10: 7444.
Takiyama M, Matsumoto T, Watanabe J. LC-MS/MS detection of Citrus unshiu peel-derived flavonoids in the plasma and brain after oral administration of yokukansankachimpihange in rats. Xenobiotica 2019; 49: 1-35.
Wang M, Zheng J, Zhong Z, Song M, Wu X. Tissue distribution of nobiletin and its metabolites in mice after oral administration of nobiletin Fed Am Soci Exp Biol 2013; 27: 1253.
Nakajima A, Ohizumi Y. Potential benefits of nobiletin, a citrus flavonoid, against Alzheimer’s disease and Parkinson’s disease. Int J Mol Sci 2019; 20: 3380.
Yabuki Y, Ohizumi Y, Yokosuka A, Mimaki Y, Fukunaga K. Nobiletin treatment improves motor and cognitive deficits seen in mptp-induced parkinson model mice. Neuroscience 2014; 259: 126-41.
Goh JXH, Tan LT, Goh JK. Nobiletin and derivatives: Functional compounds from citrus fruit peel for colon cancer chemoprevention. Cancers (Basel) 2019; 11: 867.
Shu Z, Yang B, Zhao H, Xu B, Jiao W, Wang Q. Tangeretin exerts anti-neuroinflammatory effects via NF-κB modulation in lipopolysaccharide-stimulated microglial cells. Int Immunopharmacol 2014; 19: 275-82.
Okuyama S, Fukata T, Nishigawa Y, et al. Citrus flavonoid improves MK-801-induced locomotive hyperactivity: Possible relevance to schizophrenia. J Funct Foods 2013; 5: 2002-6.
Cho J. 2006. Antioxidant and neuroprotective effects of hesperidin and its aglyconehesperetin. Arch Pharm Res 2013; 29: 699-706.
Tamilselvam K, Braidy N, Manivasagam T, Essa MM, Prasad NR, Karthikeyan S. Neuroprotective effects of hesperidin a plant flavanone on rotenone-induced oxidative stress and apoptosis in a cellular model for Parkinson’s disease. Oxid Med Cell Longev 2013; 10: 11.
Morelli S, Piscioneri A, Salerno S, Al-Fageeh MB, Drioli E, de Bartolo L. Neuroprotective effect of didymin on hydrogen peroxide-induced injury in the neuronal membrane system. Cells Tissues Organs 2014; 199: 184-200.
Vauzour D, Corona G, Spencer JPE. Caffeic acid, tyrosol and p-coumaric acid are potent inhibitors of 5-Scysteinyl-dopamine induced neurotoxicity. Arch Biochem Biophys 2010; 501: 106-11.
Ojha S, Javed H, Azimullah S. AbulKhair, S.B.; Haque, M.E. Neuroprotective potential of ferulic acid in the rotenone model of parkinson’s disease. Drug Des Devel Ther 2015; 9: 5499-510.
Zare K, Eidi A, Roghani M, Rohani AH. The neuroprotective potential of sinapic acid in the 6-hydroxydopamine-induced hemi-parkinsonian rat. Metab Brain Dis 2015; 30: 205-13.
Biondo PBF, Carbonera F, Zawadzki F, et al. Antioxidant capacity and identification of bioactive compounds by GC-MS of essential oils from spices, herbs and citrus. Curr Bioact Compd 2017; 13: 137-43.
Benavente-Garcia O, Castillo J, Alcaraz M, Vicente V, Del Rio JA, Ortuno A. Beneficial action of Citrus flavonoids on multiple cancer-related biological pathways. Curr Cancer Drug Targets 2007; 7: 795-809.
Wang X, Li S, Wei C-C, et al. Anti-inflammatory effects of polymethoxy-flavones from citrus peels: a review. J Food Bioactives 2018; 3: 76-86.
Ji X, Melman N, Jacobson KA. Interactions of flavonoids and other phytochemicals with adenosine receptors. J Med Chem 1996; 39: 781-8.
Van der Walt MM, Terre’Blanche G. Benzopyrone represents a privilege scaffold to identify novel adenosine A 1 /A 2A receptor antagonists. Bioorg Chem 2018; 77: 136-43.

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Year: 2021
Published on: 18 September, 2020
Article ID: e010621186105
Pages: 18
DOI: 10.2174/1573407216999200918182514
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