{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T03:00:08Z","timestamp":1772766008251,"version":"3.50.1"},"reference-count":52,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2017,4,28]],"date-time":"2017-04-28T00:00:00Z","timestamp":1493337600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Vaccinium myrtillus (bilberry) leaf is traditionally used in southeastern Europe for the treatment of diabetes. In the present study, the ability of bilberry leaf extracts to inhibit carbohydrate-hydrolyzing enzymes and restore glutathione concentration in Hep G2 cells subjected to glucose-induced oxidative stress was investigated. A comprehensive analysis of the antioxidant activity of two bilberry leaf extracts was performed. The aqueous extract showed excellent total antioxidant and chelating activity. Its antioxidant activity in the \u03b2-carotene-linoleic acid assay was very good, reaching the activity of the antioxidant standard BHA (93.4 \u00b1 2.3% vs. 95.1 \u00b1 2.4%, respectively). The hydroethanolic extract (ethanol\/H2O, 8:2, v\/v), on the other hand, was a better radical scavenger and Fe2+ reducing agent. Furthermore, the aqueous extract was able to efficiently increase glutathione concentration in Hep G2 cells subjected to glucose-induced oxidative stress and restore it to the levels observed in non-hyperglycaemic cells. The hydroethanolic extract strongly inhibited \u03b1-glucosidase, with the IC50 statistically equal to the antidiabetic drug acarbose (0.29 \u00b1 0.02 mg\/mL vs. 0.50 \u00b1 0.01 mg\/mL, respectively). Phytochemical analysis revealed the presence of quercetin and kaemferol derivatives, as well as chlorogenic and p-coumaric acid. The study results indicate that V. myrtillus leaf may have promising properties as a supporting therapy for diabetes.<\/jats:p>","DOI":"10.3390\/molecules22050703","type":"journal-article","created":{"date-parts":[[2017,4,28]],"date-time":"2017-04-28T11:57:04Z","timestamp":1493380624000},"page":"703","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":52,"title":["Chemical Composition, Antioxidant and \u03b1-Glucosidase-Inhibiting Activities of the Aqueous and Hydroethanolic Extracts of Vaccinium myrtillus Leaves"],"prefix":"10.3390","volume":"22","author":[{"given":"Kristina","family":"Bljaji\u0107","sequence":"first","affiliation":[{"name":"Faculty of Pharmacy and Biochemistry, University of Zagreb, A. Kova\u010di\u0107a 1, 10000 Zagreb, Croatia"}]},{"given":"Roberta","family":"Petlevski","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy and Biochemistry, University of Zagreb, A. Kova\u010di\u0107a 1, 10000 Zagreb, Croatia"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8755-9960","authenticated-orcid":false,"given":"Lovorka","family":"Vuji\u0107","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy and Biochemistry, University of Zagreb, A. Kova\u010di\u0107a 1, 10000 Zagreb, Croatia"}]},{"given":"Ana","family":"\u010ca\u010di\u0107","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy and Biochemistry, University of Zagreb, A. Kova\u010di\u0107a 1, 10000 Zagreb, Croatia"},{"name":"Faculty of Sciences and Technology, University of Algarve, Campus de Gambelas, Bd. 8, 8005-139 Faro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1968-4356","authenticated-orcid":false,"given":"Nina","family":"\u0160o\u0161tari\u0107","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy and Biochemistry, University of Zagreb, A. Kova\u010di\u0107a 1, 10000 Zagreb, Croatia"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4741-0819","authenticated-orcid":false,"given":"Jasna","family":"Jablan","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy and Biochemistry, University of Zagreb, A. Kova\u010di\u0107a 1, 10000 Zagreb, Croatia"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8057-3404","authenticated-orcid":false,"given":"Isabel","family":"Saraiva de Carvalho","sequence":"additional","affiliation":[{"name":"Faculty of Sciences and Technology, University of Algarve, Campus de Gambelas, Bd. 8, 8005-139 Faro, Portugal"}]},{"given":"Marijana","family":"Zovko Kon\u010di\u0107","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy and Biochemistry, University of Zagreb, A. Kova\u010di\u0107a 1, 10000 Zagreb, Croatia"}]}],"member":"1968","published-online":{"date-parts":[[2017,4,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Kapoor, R., and Kakkar, P. (2012). Protective Role of Morin, a Flavonoid, against High Glucose Induced Oxidative Stress Mediated Apoptosis in Primary Rat Hepatocytes. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0041663"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Han, T.S., and Lean, M.E. (2016). A clinical perspective of obesity, metabolic syndrome and cardiovascular disease. JRSM Cardiovasc. Dis., 5.","DOI":"10.1177\/2048004016633371"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"5695","DOI":"10.2174\/1381612811319320005","article-title":"Hyperglycemia-induced oxidative stress and its role in diabetes mellitus related cardiovascular diseases","volume":"19","author":"Fiorentino","year":"2013","journal-title":"Curr. Pharm. Des."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.mam.2008.08.006","article-title":"Glutathione: Overview of its protective roles, measurement, and biosynthesis","volume":"30","author":"Forman","year":"2009","journal-title":"Mol. Asp. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.mam.2008.08.003","article-title":"Glutathione in liver diseases and hepatotoxicity","volume":"30","author":"Yuan","year":"2009","journal-title":"Mol. Asp. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.1016\/S0006-2952(02)01172-3","article-title":"Cellular glutathione and thiols metabolism","volume":"64","author":"Dickinson","year":"2002","journal-title":"Biochem. Pharmacol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"942","DOI":"10.1016\/j.nutres.2013.07.011","article-title":"Olive leaf down-regulates the oxidative stress and immune dysregulation in streptozotocin-induced diabetic mice","volume":"33","author":"Park","year":"2013","journal-title":"Nutr. Res."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Lin, D., Xiao, M., Zhao, J., Li, Z., Xing, B., Li, X., Kong, M., Li, L., Zhang, Q., and Liu, Y. (2016). An Overview of Plant Phenolic Compounds and Their Importance in Human Nutrition and Management of Type 2 Diabetes. Mol. Basel Switz., 21.","DOI":"10.3390\/molecules21101374"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e973878","DOI":"10.1155\/2015\/973878","article-title":"Determination of Heavy Metals Concentration in Traditional Herbs Commonly Consumed in the United Arab Emirates","volume":"2015","author":"Dghaim","year":"2015","journal-title":"J. Environ. Publ. Health"},{"key":"ref_10","first-page":"97","article-title":"Evaluation of Heavy Metals Contents in Spices and Herbs Available on the Polish Market","volume":"16","author":"Krejpcio","year":"2007","journal-title":"Pol. J. Environ. Stud."},{"key":"ref_11","first-page":"77","article-title":"Heavy Metal Contents in Spices from Markets in Sarajevo, Bosnia and Herzegovina (Sadr\u017eaj te\u0161kih metala u za\u010dinskom bilju s tr\u017ei\u0161ta u Sarajevu, Bosna i Hercegovina)","volume":"63","author":"Badema","year":"2014","journal-title":"Kem. Ind."},{"key":"ref_12","first-page":"315","article-title":"Vaccinium myrtillus as an antidiabetic medicinal plant\u2014Research through the ages","volume":"65","author":"Schuster","year":"2010","journal-title":"Pharmacy"},{"key":"ref_13","unstructured":"Ga\u0161par, K., and Zovko Kon\u010di\u0107, M. (2013, January 3\u20136). Traditional herbal products used for the management of diabetes in Croatia. Proceedings of the International Conference on Natural Products Utilization: From Plants to Pharmacy Shelf, Bansko, Bulgaria."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Pieroni, A., and Quave, C.L. (2014). Ways the Lukomir Highlanders of Bosnia and Herzegovina Treat Diabetes. Ethnobotany and Biocultural Diversities in the Balkans, Springer.","DOI":"10.1007\/978-1-4939-1492-0"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4920","DOI":"10.1007\/s11356-012-1461-4","article-title":"A comparative study of heavy metal accumulation and antioxidant responses in Vaccinium myrtillus L. leaves in polluted and non-polluted areas","volume":"20","author":"Barczyk","year":"2013","journal-title":"Environ. Sci. Pollut. Res. Int."},{"key":"ref_16","unstructured":"Wichtl, M. (2004). Myrtilli folium. Herbal Drugs and Phytopharmaceuticals, Medpharm Stuttgart. [3rd ed.]."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/S0048-9697(00)00890-1","article-title":"Comparison of the element composition in several plant species and their substrate from a 1,500,000-km2 area in Northern Europe","volume":"278","author":"Reimann","year":"2001","journal-title":"Sci. Total Environ."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.jep.2016.12.006","article-title":"In vitro studies to evaluate the wound healing properties of Calendula officinalis extracts","volume":"196","author":"Nicolaus","year":"2016","journal-title":"J. Ethnopharmacol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1002\/pca.2462","article-title":"Phenolic distribution in liquid preparations of Vaccinium myrtillus L. and Vaccinium vitis idaea L.","volume":"24","author":"Ieri","year":"2013","journal-title":"Phytochem. Anal. PCA"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"12015","DOI":"10.1021\/jf503521m","article-title":"Characterization of metabolite profiles of leaves of bilberry (Vaccinium myrtillus L.) and lingonberry (Vaccinium vitis-idaea L.)","volume":"62","author":"Liu","year":"2014","journal-title":"J. Agric. Food Chem."},{"key":"ref_21","first-page":"59","article-title":"Morphological characteristics of Vaccinium \u00d7 intermedium Ruthe","volume":"51","author":"Ponikierska","year":"2004","journal-title":"Dendrobiol. Pol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.1016\/j.foodchem.2010.07.032","article-title":"Identification and quantification of flavonol aglycons in cactus pear (Opuntia ficus indica) fruit using a commercial pectinase and cellulase preparation","volume":"124","author":"Kroh","year":"2011","journal-title":"Food Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1111\/j.1541-4337.2011.00173.x","article-title":"Phenol-Based Antioxidants and the In Vitro Methods Used for Their Assessment","volume":"11","author":"Craft","year":"2012","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Pokorny, J., Yanishlieva, N., and Gordon, M.H. (2001). Antioxidants in Food: Practical Applications, CRC Press.","DOI":"10.1201\/9781439823057"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1006\/abio.1999.4019","article-title":"Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: Specific application to the determination of vitamin E","volume":"269","author":"Prieto","year":"1999","journal-title":"Anal. Biochem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"940","DOI":"10.1016\/j.foodchem.2007.04.038","article-title":"Antioxidant activity and phenolic compounds in 32 selected herbs","volume":"105","author":"Czemerys","year":"2007","journal-title":"Food Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1515\/jbcpp-2013-0141","article-title":"Caffeic and chlorogenic acids inhibit key enzymes linked to type 2 diabetes (in vitro): A comparative study","volume":"26","author":"Oboh","year":"2015","journal-title":"J. Basic Clin. Physiol. Pharmacol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1016\/j.fitote.2009.08.019","article-title":"Antioxidant and enzyme inhibition activities and chemical profiles of Polygonum sachalinensis F.Schmidt ex Maxim (Polygonaceae)","volume":"81","author":"Fan","year":"2010","journal-title":"Fitoterapia"},{"key":"ref_29","first-page":"763","article-title":"Hypoglycemic activity of Pyrus biossieriana Buhse leaf extract and arbutin: Inhibitory effects on alpha amylase and alpha glucosidase","volume":"4","author":"Yousefi","year":"2013","journal-title":"Casp. J. Intern. Med."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1472-6882-12-110","article-title":"In vitro inhibitory effects of plant-based foods and their combinations on intestinal \u03b1-glucosidase and pancreatic \u03b1-amylase","volume":"12","author":"Adisakwattana","year":"2012","journal-title":"BMC Complement. Altern. Med."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"11463","DOI":"10.1021\/jf903083h","article-title":"Comparative Evaluation of Quercetin, Isoquercetin and Rutin as Inhibitors of \u03b1-Glucosidase","volume":"57","author":"Li","year":"2009","journal-title":"J. Agric. Food Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1016\/j.tiv.2009.10.020","article-title":"Apoptosis in HepG2 cells exposed to high glucose","volume":"24","author":"Chandrasekaran","year":"2010","journal-title":"Toxicol. In Vitro"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.2174\/1389450115666141024113925","article-title":"Pharmacological applications of antioxidants: Lights and shadows","volume":"15","author":"Saso","year":"2014","journal-title":"Curr. Drug Targets"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1428","DOI":"10.3390\/jcm4071428","article-title":"Mitochondrial Glutathione in Diabetic Nephropathy","volume":"4","author":"Lash","year":"2015","journal-title":"J. Clin. Med."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"20023","DOI":"10.3390\/molecules191220023","article-title":"Effects of Selected Dietary Secondary Metabolites on Reactive Oxygen Species Production Caused by Iron (II) Autoxidation","volume":"19","author":"Chobot","year":"2014","journal-title":"Molecules"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1089\/jmf.2011.1921","article-title":"Pretreatment with the Total Flavone Glycosides of Flos Abelmoschus manihot and Hyperoside Prevents Glomerular Podocyte Apoptosis in Streptozotocin-Induced Diabetic Nephropathy","volume":"15","author":"Zhou","year":"2012","journal-title":"J. Med. Food"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"740","DOI":"10.1007\/s11418-016-1007-z","article-title":"Hyperoside reduces albuminuria in diabetic nephropathy at the early stage through ameliorating renal damage and podocyte injury","volume":"70","author":"Zhang","year":"2016","journal-title":"J. Nat. Med."},{"key":"ref_38","first-page":"65","article-title":"Pharmacological evaluation of hyperin for antihyperglycemic activity and effect on lipid profile in diabetic rats","volume":"51","author":"Verma","year":"2013","journal-title":"Indian J. Exp. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1155\/2013\/297154","article-title":"Roles of Chlorogenic Acid on Regulating Glucose and Lipids Metabolism: A Review","volume":"2013","author":"Meng","year":"2013","journal-title":"Evid. Based Complement. Altern. Med."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1345","DOI":"10.1007\/s00394-016-1206-0","article-title":"Coffee consumption, obesity and type 2 diabetes: A mini-review","volume":"55","author":"Santos","year":"2016","journal-title":"Eur. J. Nutr."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"989","DOI":"10.1007\/s12272-016-0771-3","article-title":"The attenuation of chlorogenic acid on oxidative stress for renal injury in streptozotocin-induced diabetic nephropathy rats","volume":"39","author":"Ye","year":"2016","journal-title":"Arch. Pharm. Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1016\/S0076-6879(99)99017-1","article-title":"Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent","volume":"Volume 299","author":"Singleton","year":"1999","journal-title":"Methods in Enzymology"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1016\/S0308-8146(03)00216-4","article-title":"Antioxidant activity of propolis of various geographic origins","volume":"84","author":"Kumazawa","year":"2004","journal-title":"Food Chem."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Ermer, J. (2005). Method Validation in Pharmaceutical Analysis: A Guide to Best Practice, Wiley-VCH.","DOI":"10.1002\/3527604685"},{"key":"ref_45","unstructured":"(2013). European Pharmacopoeia, Council of Europe. [8th ed.]."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"2176","DOI":"10.1016\/j.fct.2010.05.025","article-title":"Evaluation of antioxidant activities and phenolic content of Berberis vulgaris L. and Berberis croatica Horvat","volume":"48","author":"Kremer","year":"2010","journal-title":"Food Chem. Toxicol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1006\/abio.1996.0292","article-title":"The ferric reducing ability of plasma (FRAP) as a measure of \u201cantioxidant power\u201d: The FRAP assay","volume":"239","author":"Benzie","year":"1996","journal-title":"Anal. Biochem."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"6232","DOI":"10.3390\/molecules16086232","article-title":"Antiradical, Chelating and Antioxidant Activities of Hydroxamic Acids and Hydroxyureas","volume":"16","author":"Zorc","year":"2011","journal-title":"Molecules"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1231","DOI":"10.1016\/S0891-5849(98)00315-3","article-title":"Antioxidant activity applying an improved ABTS radical cation decolorization assay","volume":"26","author":"Re","year":"1999","journal-title":"Free Radic. Biol. Med."},{"key":"ref_50","first-page":"915","article-title":"Identification of proglycemic and antihyperglycemic activity in antioxidant rich fraction of some common food grains","volume":"18","author":"Tiwari","year":"2011","journal-title":"Int. Food Res. J."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/j.ifset.2006.06.001","article-title":"Inhibitory potential of herb, fruit, and fungal-enriched cheese against key enzymes linked to type 2 diabetes and hypertension","volume":"8","author":"Apostolidis","year":"2007","journal-title":"Innov. Food Sci. Emerg. Technol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1051","DOI":"10.1016\/j.lfs.2003.07.020","article-title":"Protective effect of acteoside on carbon tetrachloride-induced hepatotoxicity","volume":"74","author":"Lee","year":"2004","journal-title":"Life Sci."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/22\/5\/703\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T18:34:06Z","timestamp":1760207646000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/22\/5\/703"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,4,28]]},"references-count":52,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2017,5]]}},"alternative-id":["molecules22050703"],"URL":"https:\/\/doi.org\/10.3390\/molecules22050703","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,4,28]]}}}