{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T04:33:34Z","timestamp":1760243614491,"version":"build-2065373602"},"reference-count":58,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2013,10,28]],"date-time":"2013-10-28T00:00:00Z","timestamp":1382918400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antioxidants"],"abstract":"<jats:p>Plant phytochemicals are described as possessing considerable neuroprotective properties, due to radical scavenging capacity and acetylcholinesterase inhibitory activity, important bioactivities in neurodegeneration. Antirrhinum lopesianum is a rare endemism from the Iberian Peninsula, occurring at the northeastern border between Portugal and Spain. It is classified as Endangered, due to its highly fragmented geographical occupation, facing a high risk of extinction in the Portuguese territory, within 20 years. Here, we describe for the first time the chemical characterization of extracts of the species concerning total phenol content, flavonoid content and antioxidant properties. The profile of high performance liquid chromatography with diode array detector (HPLC-DAD) of the polyphenol-enriched fraction of plant extracts was also performed, showing the great potential of the species as a source of bioactive phytochemical compounds. A. lopesianum\u2019s potential for neuroprotection was revealed by a significant acetylcholinesterase inhibitory activity and also by a neuroprotective effect on a human cell model of neurodegeneration. Moreover, this is the first report describing a successful procedure for the in vitro propagation of this endangered species. The comparison of phenolic content and the HPLC-DAD profile of wild and in vitro propagated plants revealed that in vitro plants maintain the ability to produce secondary metabolites, but the profiles are differentially affected by the growth regulators. The results presented here greatly contribute to the value for this species regarding its potential as a source of phytochemicals with prospective neuroprotective health benefits.<\/jats:p>","DOI":"10.3390\/antiox2040273","type":"journal-article","created":{"date-parts":[[2013,10,28]],"date-time":"2013-10-28T12:00:27Z","timestamp":1382961627000},"page":"273-292","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Valuing the Endangered Species Antirrhinum lopesianum: Neuroprotective Activities and Strategies for in vitro Plant Propagation"],"prefix":"10.3390","volume":"2","author":[{"given":"Andreia","family":"Gomes","sequence":"first","affiliation":[{"name":"Institute of Chemical and Biological Technology, New University of Lisbon, Av. da Rep\u00fablica, Oeiras 2780-157, Portugal"},{"name":"Institute of Experimental and Technological Biology, Apartado 12, Oeiras 2781-901, Portugal"}]},{"given":"Sofia","family":"Fortalezas","sequence":"additional","affiliation":[{"name":"Institute of Chemical and Biological Technology, New University of Lisbon, Av. da Rep\u00fablica, Oeiras 2780-157, Portugal"}]},{"given":"Rui","family":"Pimp\u00e3o","sequence":"additional","affiliation":[{"name":"Institute of Chemical and Biological Technology, New University of Lisbon, Av. da Rep\u00fablica, Oeiras 2780-157, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4639-6883","authenticated-orcid":false,"given":"In\u00eas","family":"Figueira","sequence":"additional","affiliation":[{"name":"Institute of Chemical and Biological Technology, New University of Lisbon, Av. da Rep\u00fablica, Oeiras 2780-157, Portugal"}]},{"given":"Jo\u00e3o","family":"Maroco","sequence":"additional","affiliation":[{"name":"University Institute of Psychological, Social and Life Sciences, Rua Jardim do Tabaco 34,  Lisboa 1149-051, Portugal"}]},{"given":"Carlos","family":"Aguiar","sequence":"additional","affiliation":[{"name":"Mountain Research Center, Polytechnic Institute of Bragan\u00e7a, Apartado 1172, Bragan\u00e7a 5301-855, Portugal"}]},{"given":"Ricardo","family":"Ferreira","sequence":"additional","affiliation":[{"name":"Institute of Chemical and Biological Technology, New University of Lisbon, Av. da Rep\u00fablica, Oeiras 2780-157, Portugal"},{"name":"Department of Botany and Biological Engineering, Institute of Agronomy, Technical University of Lisbon, Tapada da Ajuda, Lisboa 1349-017, Portugal"}]},{"given":"C\u00e9lia","family":"Miguel","sequence":"additional","affiliation":[{"name":"Institute of Chemical and Biological Technology, New University of Lisbon, Av. da Rep\u00fablica, Oeiras 2780-157, Portugal"},{"name":"Institute of Experimental and Technological Biology, Apartado 12, Oeiras 2781-901, Portugal"}]},{"given":"Cl\u00e1udia","family":"Santos","sequence":"additional","affiliation":[{"name":"Institute of Chemical and Biological Technology, New University of Lisbon, Av. da Rep\u00fablica, Oeiras 2780-157, Portugal"},{"name":"Institute of Experimental and Technological Biology, Apartado 12, Oeiras 2781-901, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2013,10,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1022","DOI":"10.1021\/np030096l","article-title":"Natural products as sources of new drugs over the period 1981\u20132002","volume":"66","author":"Newman","year":"2003","journal-title":"J. Nat. Prod."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1002\/ptr.3258","article-title":"Metabolomics for bioactivity assessment of natural products","volume":"25","author":"Yuliana","year":"2011","journal-title":"Phytother. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/j.drudis.2007.10.010","article-title":"Plant-derived compounds in clinical trials","volume":"13","author":"Saklani","year":"2008","journal-title":"Drug Discov. Today"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1100\/tsw.2010.44","article-title":"Cucurbitacin: Ancient compound shedding new light on cancer treatment","volume":"10","author":"Lee","year":"2010","journal-title":"Sci. World J."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1021\/np068054v","article-title":"Natural products as sources of new drugs over the last 25 years","volume":"70","author":"Newman","year":"2007","journal-title":"J. Nat. Prod."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1002\/mnfr.200500156","article-title":"Green tea catechins as brain-permeable, natural iron chelators-antioxidants for the treatment of neurodegenerative disorders","volume":"50","author":"Mandel","year":"2006","journal-title":"Mol. Nutr. Food. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.ejphar.2006.06.025","article-title":"Emerging role of polyphenolic compounds in the treatment of neurodegenerative diseases: A review of their intracellular targets","volume":"545","author":"Ramassamy","year":"2006","journal-title":"Eur. J. Pharmacol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1039\/C0NP00027B","article-title":"Natural products as a source of Alzheimer\u2019s drug leads","volume":"28","author":"Williams","year":"2011","journal-title":"Nat. Prod. Rep."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1403","DOI":"10.1017\/S1368980009992990","article-title":"Flavonoid intake and disability-adjusted life years due to Alzheimer\u2019s and related dementias: A population-based study involving twenty-three developed countries","volume":"13","author":"Beking","year":"2010","journal-title":"Public Health Nutr."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1002\/gps.2610","article-title":"The economic impact of dementia in Europe in 2008-cost estimates from the Eurocode project","volume":"26","author":"Wimo","year":"2011","journal-title":"Int. J. Geriatr. Psychiatry"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"S18","DOI":"10.1038\/nrn1434","article-title":"Oxidative stress in neurodegeneration: Cause or consequence?","volume":"10","author":"Andersen","year":"2004","journal-title":"Nat. Med."},{"key":"ref_12","first-page":"48","article-title":"Medicinal chemistry approaches for the treatment and prevention of Alzheimer\u2019s disease","volume":"23","author":"Bachurin","year":"2003","journal-title":"Med. Chem. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"690","DOI":"10.1136\/pgmj.2004.018903","article-title":"Update on myasthenia gravis","volume":"80","author":"Thanvi","year":"2004","journal-title":"Postgrad. Med. J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1038\/35067589","article-title":"Acetylcholinesterase\u2014New roles for an old actor","volume":"2","author":"Soreq","year":"2001","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1016\/j.phymed.2007.02.002","article-title":"Acetylcholinesterase inhibitors from plants","volume":"14","author":"Mukherjee","year":"2007","journal-title":"Phytomedicine"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"465","DOI":"10.2165\/00002018-199819060-00004","article-title":"Cholinesterase inhibitors in the treatment of Alzheimer\u2019s disease: A comparison of tolerability and pharmacology","volume":"19","author":"Nordberg","year":"1998","journal-title":"Drug Saf."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1475","DOI":"10.1016\/S0891-5849(02)01113-9","article-title":"Is oxidative damage the fundamental pathogenic mechanism of Alzheimer\u2019s and other neurodegenerative diseases?","volume":"33","author":"Perry","year":"2002","journal-title":"Free Radic. Biol. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1297","DOI":"10.1111\/j.1365-2699.2008.02059.x","article-title":"A geographical pattern of Antirrhinum (Scrophulariaceae) speciation since the Pliocene based on plastid and nuclear DNA polymorphisms","volume":"36","author":"Vargas","year":"2009","journal-title":"J. Biogeogr."},{"key":"ref_19","unstructured":"Amich Garc\u00eda, F., Bernardos Hern\u00e1ndez, S., Gonz\u00e1lez Talav\u00e1n, A., Barreto Caldas, F., and Alves, P. IUCN Red List of Threatened Species, Version 2013.1. >Antirrhinum lopesianum. Available online:http:\/\/www.iucnredlist.org."},{"key":"ref_20","first-page":"231","article-title":"Antirrhinum lopesianum Rothm., novedad para la flora espa\u00f1ola","volume":"62","author":"Amich","year":"1989","journal-title":"Bol. Soc. Brot."},{"key":"ref_21","first-page":"129","article-title":"Apontamentos para o estudo da flora transmontana","volume":"2","author":"Coutinho","year":"1883","journal-title":"Bol. Soc. Brot."},{"key":"ref_22","first-page":"234","article-title":"A flora do concelho de Vimioso e arredores","volume":"4","author":"Lopes","year":"1926","journal-title":"Bol. Soc. Brot."},{"key":"ref_23","unstructured":"Rothmaler, W. (1956). Taxonomische Monographie der Gattung Antirrhinum, Akademie Verlag."},{"key":"ref_24","unstructured":"Aguiar, C. (1996). Distribui\u00e7\u00e3o Geogr\u00e1fica e Estatuto de Amea\u00e7a das esp\u00e9cies da Flora a Proteger, ESAB."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1080\/11263500500499692","article-title":"Conservation status of the threatened Iberian Peninsula narrow endemic Antirrhinum lopesianum Rothm. (Scrophulariaceae)","volume":"140","author":"Bernardos","year":"2006","journal-title":"Plant Biosyst."},{"key":"ref_26","unstructured":"(1992). Directive 92\/43 of the Council of the European Community on the Conservation of Habitats and Wild Fauna and Flora, European Community."},{"key":"ref_27","unstructured":"Bilz, M., Kell, S.P., Maxted, N., and Lansdown, R.V. (2011). European Red List of Vascular Plants, Publications Office of the European Union."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1059","DOI":"10.1007\/s10531-004-8413-3","article-title":"In vitro micropropagation of endangered Rhododendron ponticum L. subsp. baeticum (Boissier & Reuter) Handel-Mazzetti","volume":"14","author":"Almeida","year":"2005","journal-title":"Biodivers. Conserv."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1007\/s00299-006-0230-6","article-title":"In vitro propagation of an endangered medicinal plant Saussurea involucrata Kar. et Kir.","volume":"26","author":"Guo","year":"2007","journal-title":"Plant Cell Rep."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1023\/B:BIOP.0000041084.77832.11","article-title":"Micropropagation of an endangered orchid Anoectochilus formosanus","volume":"48","author":"Ket","year":"2004","journal-title":"Biol. Plant."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"674","DOI":"10.1007\/s11627-011-9377-0","article-title":"In vitro micropropagation of three rare, endangered, and endemic rhododendron species of Northeast India","volume":"47","author":"Mao","year":"2011","journal-title":"In Vitro Cell Dev.Biol. Plant"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1007\/s10531-010-9949-z","article-title":"Crambe tataria: Actions for ex situ conservation","volume":"20","author":"Piovan","year":"2011","journal-title":"Biodivers. Conserv."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1007\/s10535-012-0280-2","article-title":"In vitro regeneration of Aristolochia tagala and production of artificial seeds","volume":"57","author":"Remya","year":"2013","journal-title":"Biol. Plant."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3","DOI":"10.3233\/BR-2010-001","article-title":"Antioxidant and antiproliferative properties of strawberry tree tissues","volume":"1","author":"Tavares","year":"2010","journal-title":"J. Berry Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"214","DOI":"10.3390\/nu2020214","article-title":"Antioxidant properties and neuroprotective capacity of strawberry tree fruit (Arbutus unedo)","volume":"2","author":"Fortalezas","year":"2010","journal-title":"Nutrients"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2576","DOI":"10.3390\/molecules15042576","article-title":"Antioxidant capacity of Macaronesian traditional medicinal plants","volume":"15","author":"Tavares","year":"2010","journal-title":"Molecules"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1016\/0006-2952(61)90145-9","article-title":"A new and rapid colorimetric determination of acetylcholinesterase activity","volume":"7","author":"Ellman","year":"1961","journal-title":"Biochem. Pharmacol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"239","DOI":"10.3109\/13880209.2011.596209","article-title":"Bioactive compounds from endemic plants of Southwest Portugal: Inhibition of acetylcholinesterase and radical scavenging activities","volume":"50","author":"Tavares","year":"2011","journal-title":"Pharma Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"562","DOI":"10.1016\/j.foodchem.2012.05.023","article-title":"The neuroprotective potential of phenolic-enriched fractions from four Juniperus species found in Portugal","volume":"135","author":"Tavares","year":"2012","journal-title":"Food Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1111\/j.1399-3054.1962.tb08052.x","article-title":"A revised medium for rapid growth and bio assays with tobacco tissue cultures","volume":"15","author":"Murashige","year":"1962","journal-title":"Physiol. Plant"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1085","DOI":"10.2307\/3870059","article-title":"Stress-induced phenylpropanoid metabolism","volume":"7","author":"Dixon","year":"1995","journal-title":"Plant Cell"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1016\/j.plaphy.2005.01.020","article-title":"Effect of water and temperature stress on the content of active constituents of Hypericum brasiliense choisy","volume":"43","author":"Mazzafera","year":"2005","journal-title":"Plant Physiol. Biochem."},{"key":"ref_43","first-page":"417","article-title":"The effects of flooding and drought stresses on the antioxidant constituents in sweet potato leaves","volume":"47","author":"Lin","year":"2006","journal-title":"Bot. Stud."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1016\/j.phytochem.2006.10.014","article-title":"Antiproliferative activity is predominantly associated with ellagitannins in raspberry extracts","volume":"68","author":"Ross","year":"2007","journal-title":"Phytochemistry"},{"key":"ref_45","first-page":"250","article-title":"A review on pharmacological studies of Bacopa monniera","volume":"1","author":"Bammidi","year":"2011","journal-title":"J. Chem. Biol. Phys. Sci."},{"key":"ref_46","first-page":"637","article-title":"Constituents of Plantago major subsp. intermedia with antioxidant and anticholinesterase capacities","volume":"35","author":"Kolak","year":"2011","journal-title":"Turk. J. Chem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/S1016-8478(23)17016-6","article-title":"Inhibitory effect of ursolic acid purified from Origanum majorana L. on the acetylcholinesterase","volume":"11","author":"Chung","year":"2001","journal-title":"Mol. Cells"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1016\/S0367-326X(02)00314-3","article-title":"Bioactive constituents of Leptadenia arborea","volume":"74","author":"Hamed","year":"2003","journal-title":"Fitoterapia"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1007\/s12263-011-0255-5","article-title":"Do polyphenols enter the brain and does it matter? Some theoretical and practical considerations","volume":"7","author":"Schaffer","year":"2012","journal-title":"Genes Nutr."},{"key":"ref_50","unstructured":"(1994). Genebank Standards, Food and Agriculture Organization of the United Nations, International Plant Genetic Resources Institute."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1007\/BF00042282","article-title":"Regeneration of plants from Antirrhinum majus L. callus","volume":"17","author":"Atkinson","year":"1989","journal-title":"Plant Cell Tissue Org."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1080\/14620316.1996.11515377","article-title":"Micropropagation of commercial and wild genotypes of snapdragon (Antirrhinum spp.)","volume":"71","author":"Tapia","year":"1996","journal-title":"J. Hortic. Sci."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"661","DOI":"10.17660\/ActaHortic.1987.212.111","article-title":"In vitro culture of shoot-tip meristems in some higher plants","volume":"212","author":"Sangwan","year":"1987","journal-title":"Acta Hortic. (ISHS)"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1111\/j.1399-3054.1978.tb08633.x","article-title":"Induction of adventitious buds on embryos of Norway spruce grown in vitro","volume":"44","author":"Arnold","year":"1978","journal-title":"Physiol. Plant"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1325","DOI":"10.1007\/s00299-010-0919-4","article-title":"Quality evaluation of snow lotus (Saussurea): quantitative chemical analysis and antioxidant activity assessment","volume":"29","author":"Qiu","year":"2010","journal-title":"Plant Cell Rep."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"422","DOI":"10.1007\/s11627-010-9299-2","article-title":"In vitro culture of Hypericum rumeliacum Boiss. and production of phenolics and flavonoids","volume":"46","author":"Danova","year":"2010","journal-title":"In Vitro Cell Dev. Biol. Plant"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"11R","DOI":"10.1039\/a606499j","article-title":"Analytical chemistry of fruit bioflavonoids\u2014A review","volume":"122","author":"Robards","year":"1997","journal-title":"Analyst"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"15118","DOI":"10.1021\/jp9052538","article-title":"Absorption and emission of the apigenin and luteolin flavonoids: A TDDFT investigation","volume":"113","author":"Amat","year":"2009","journal-title":"J. Phys. Chem. A"}],"container-title":["Antioxidants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-3921\/2\/4\/273\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T21:50:12Z","timestamp":1760219412000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-3921\/2\/4\/273"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,10,28]]},"references-count":58,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2013,12]]}},"alternative-id":["antiox2040273"],"URL":"https:\/\/doi.org\/10.3390\/antiox2040273","relation":{},"ISSN":["2076-3921"],"issn-type":[{"type":"electronic","value":"2076-3921"}],"subject":[],"published":{"date-parts":[[2013,10,28]]}}}