{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T21:47:25Z","timestamp":1784238445331,"version":"3.55.0"},"reference-count":34,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,2,6]],"date-time":"2026-02-06T00:00:00Z","timestamp":1770336000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Science, Technological Development and Innovation of the Republic of Serbia","award":["451-03-137\/2025-03\/200113"],"award-info":[{"award-number":["451-03-137\/2025-03\/200113"]}]},{"name":"Ministry of Science, Technological Development and Innovation of the Republic of Serbia","award":["451-03-136\/2025-03\/200113"],"award-info":[{"award-number":["451-03-136\/2025-03\/200113"]}]},{"name":"Ministry of Science, Technological Development and Innovation of the Republic of Serbia","award":["451-03-136\/2025-03\/200003"],"award-info":[{"award-number":["451-03-136\/2025-03\/200003"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>Bilberry (Vacciniummyrtillus L., Ericaceae) is chiefly valued as an edible plant for its berries, widely consumed as a functional food, whereas the leaves, as agro-waste, remain an underutilized natural source of bioactives. The traditional use of V. myrtillus leaves is well documented, particularly for managing diabetes and gastrointestinal disorders. However, their potential spasmolytic activity, which could support such uses, remains unexplored. The main objective of this study was to evaluate the spasmolytic potential of V. myrtillus leaf extract on the gastrointestinal tract and to elucidate its underlying mechanism of action. The spray-dried 50% hydroethanolic extract of V. myrtillus leaves, obtained by double percolation, was analyzed using HPLC-DAD. The analysis revealed phenolic acids, with chlorogenic acid as the major compound, and flavonoids, predominantly isoquercitrin. Spasmolytic activity was tested on isolated rat ileum, and the mechanism of action was monitored using models of spontaneous contractions and acetylcholine-, histamine-, CaCl2\u2212, Bay K8644-, L-NAME-, ODQ-, apamin-, BaCl2\u2212, charybdotoxin-, glibenclamide-, TRAM-34-, and quinine-modified contractions. The extract\u2019s activity on isolated ileum strips is primarily mediated via Ca2+ channels, cGMP, histamine, and NO pathways. Overall, this study affirms V. myrtillus leaves as a valuable source of phenolic compounds with potential for treating spasmodic gastrointestinal disorders.<\/jats:p>","DOI":"10.3390\/plants15030504","type":"journal-article","created":{"date-parts":[[2026,2,6]],"date-time":"2026-02-06T11:10:38Z","timestamp":1770376238000},"page":"504","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["From Agro-Waste to Therapeutic Potential: Spasmolytic Mechanisms of Vaccinium myrtillus L. Leaf Extract on Isolated Rat Ileum"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-1440-9257","authenticated-orcid":false,"given":"Nemanja","family":"Kiti\u0107","sequence":"first","affiliation":[{"name":"Department of Physiology, Faculty of Medicine, University of Ni\u0161, Ave. Dr. Zorana \u0110in\u0111i\u0107a 81, 18000 Ni\u0161, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2086-9593","authenticated-orcid":false,"given":"Katarina","family":"\u0160avikin","sequence":"additional","affiliation":[{"name":"Institute for Medicinal Plants Research \u201cDr. Josif Pan\u010di\u0107\u201d, Tadeu\u0161a Ko\u0161\u0107u\u0161ka 1, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8551-274X","authenticated-orcid":false,"given":"Du\u0161anka","family":"Kiti\u0107","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Faculty of Medicine, University of Ni\u0161, Ave. Dr. Zorana \u0110in\u0111i\u0107a 81, 18000 Ni\u0161, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0039-0929","authenticated-orcid":false,"given":"Milo\u0161","family":"Jovanovi\u0107","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Faculty of Medicine, University of Ni\u0161, Ave. Dr. Zorana \u0110in\u0111i\u0107a 81, 18000 Ni\u0161, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6951-9957","authenticated-orcid":false,"given":"Milica","family":"Randjelovi\u0107","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Faculty of Medicine, University of Ni\u0161, Ave. Dr. Zorana \u0110in\u0111i\u0107a 81, 18000 Ni\u0161, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9684-2637","authenticated-orcid":false,"given":"Jelena","family":"\u017divkovi\u0107","sequence":"additional","affiliation":[{"name":"Institute for Medicinal Plants Research \u201cDr. Josif Pan\u010di\u0107\u201d, Tadeu\u0161a Ko\u0161\u0107u\u0161ka 1, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2880-6519","authenticated-orcid":false,"given":"Bojana","family":"Miladinovi\u0107","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Faculty of Medicine, University of Ni\u0161, Ave. Dr. Zorana \u0110in\u0111i\u0107a 81, 18000 Ni\u0161, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6736-3637","authenticated-orcid":false,"given":"Nada","family":"\u0106uji\u0107 Nikoli\u0107","sequence":"additional","affiliation":[{"name":"Institute for Medicinal Plants Research \u201cDr. Josif Pan\u010di\u0107\u201d, Tadeu\u0161a Ko\u0161\u0107u\u0161ka 1, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8064-4388","authenticated-orcid":false,"given":"Nenad","family":"Stojiljkovi\u0107","sequence":"additional","affiliation":[{"name":"Department of Physiology, Faculty of Medicine, University of Ni\u0161, Ave. Dr. Zorana \u0110in\u0111i\u0107a 81, 18000 Ni\u0161, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-1814-9844","authenticated-orcid":false,"given":"Suzana","family":"Brankovi\u0107","sequence":"additional","affiliation":[{"name":"Department of Physiology, Faculty of Medicine, University of Ni\u0161, Ave. Dr. Zorana \u0110in\u0111i\u0107a 81, 18000 Ni\u0161, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Tundis, R., Tenuta, M.C., Loizzo, M.R., Bonesi, M., Finetti, F., Trabalzini, L., and Deguin, B. (2021). Vaccinium species (Ericaceae): From chemical composition to bio-functional activities. Appl. Sci., 11.","DOI":"10.3390\/app11125655"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1016\/j.foodchem.2016.06.042","article-title":"Seasonal variations of the phenolic constituents in bilberry (Vaccinium myrtillus L.) leaves, stems and fruits, and their antioxidant activity","volume":"213","author":"Bujor","year":"2016","journal-title":"Food Chem."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"\u0218tef\u0103nescu, R., Laczk\u00f3-Z\u00f6ld, E., \u0150sz, B.E., and Vari, C.E. (2022). An updated systematic review of Vaccinium myrtillus leaves: Phytochemistry and pharmacology. Pharmaceutics, 15.","DOI":"10.3390\/pharmaceutics15010016"},{"key":"ref_4","unstructured":"EMA Herbal Medicinal Products (2025, December 17). Assessment Report on Vaccinium myrtillus L., Fructus Recens and Vaccinium myrtillus L., Fructus Siccus. Available online: https:\/\/www.ema.europa.eu\/en\/documents\/herbal-report\/final-assessment-report-vaccinium-myrtillus-l-fructus-recens-and-vaccinium-myrtillus-l-fructus-siccus_en.pdf."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3240","DOI":"10.1038\/s41598-021-82207-2","article-title":"Assessment of cytotoxicity and antioxidant properties of berry leaves as by-products with potential application in cosmetic and pharmaceutical products","volume":"11","author":"Ziemlewska","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Elferjane, M.R., Milutinovi\u0107, V., Jovanovi\u0107 Krivoku\u0107a, M., Taherzadeh, M.J., Pietrzak, W., Marinkovi\u0107, A., and Jovanovi\u0107, A.A. (2024). Vaccinium myrtillus L. leaf waste as a source of biologically potent compounds: Optimization of polyphenol extractions, chemical profile, and biological properties of the extracts. Pharmaceutics, 16.","DOI":"10.3390\/pharmaceutics16060740"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Ferlemi, A.-V., and Lamari, F.N. (2016). Berry leaves: An alternative source of bioactive natural products of nutritional and medicinal value. Antioxidants, 5.","DOI":"10.3390\/antiox5020017"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Vrancheva, R., Ivanov, I., Dincheva, I., Badjakov, I., and Pavlov, A. (2021). Triterpenoids and other non-polar compounds in leaves of wild and cultivated Vaccinium species. Plants, 10.","DOI":"10.3390\/plants10010094"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"736","DOI":"10.1016\/j.jff.2015.02.041","article-title":"Comparison of phenolic compounds and antioxidant potential between selected edible fruits and their leaves","volume":"14","author":"Teleszko","year":"2015","journal-title":"J. Funct. Foods"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"577","DOI":"10.9755\/ejfa.v26i7.16404","article-title":"Vaccinium (Ericaceae): Ethnobotany and pharmacological potentials","volume":"26","author":"Abreu","year":"2014","journal-title":"Emir. J. Food Agric."},{"key":"ref_11","first-page":"315","article-title":"Vaccinium myrtillus as an antidiabetic medicinal plant\u2013research through the ages","volume":"65","author":"Schuster","year":"2010","journal-title":"Die Pharm."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Czigle, S., Bittner Fialova, S., T\u00f3th, J., Mu\u010daji, P., Nagy, M. (2022). Treatment of gastrointestinal disorders\u2014Plants and potential mechanisms of action of their constituents. Molecules, 27.","DOI":"10.3390\/molecules27092881"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4889719","DOI":"10.1155\/2021\/4889719","article-title":"Antispasmodic potential of medicinal plants: A comprehensive review","volume":"2021","author":"Rauf","year":"2021","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_14","first-page":"3819714","article-title":"Natural antispasmodics: Source, stereochemical configuration, and biological activity","volume":"2018","author":"Bach","year":"2018","journal-title":"Biomed Res. Int."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Brasanac-Vukanovi\u0107, S., Mutic, J., Stankovic, D.M., Arsic, I., Blagojevic, N., Vukasinovic-Pesic, V., and Tadic, V.M. (2018). Wild bilberry (Vaccinium myrtillus L., Ericaceae) from Montenegro as a source of antioxidants for use in the production of nutraceuticals. Molecules, 23.","DOI":"10.3390\/molecules23081864"},{"key":"ref_16","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_17","doi-asserted-by":"crossref","unstructured":"Kuzmanovi\u0107 Nedeljkovi\u0107, S., Radan, M., \u0106uji\u0107 Nikoli\u0107, N., Mutavski, Z., Krgovi\u0107, N., Markovi\u0107, S., and \u0160avikin, K. (2023). Microencapsulated bilberry and chokeberry leaf extracts with potential health benefits. Plants, 12.","DOI":"10.3390\/plants12233979"},{"key":"ref_18","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."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Rice-Evans, C.A., and Packer, L. (2003). Flavonoids in Health and Disease, Taylor & Francis. [2nd ed.].","DOI":"10.1201\/9780367803681"},{"key":"ref_20","unstructured":"Kuhn, M.A., and Winston, D. (2012). Winston & Kuhn\u2019s Herbal Therapy and Supplements: A Scientific and Traditional Approach, Lippincott Williams & Wilkins. [2nd ed.]."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Heghes, S.C., Vostinaru, O., Rus, L.M., Mogosan, C., Iuga, C.A., and Filip, L. (2019). Antispasmodic effect of essential oils and their constituents: A review. Molecules, 24.","DOI":"10.3390\/molecules24091675"},{"key":"ref_22","first-page":"402","article-title":"In vitro antispasmodic and antiinflammatory effects of Drosera rotundifolia","volume":"54","author":"Krenn","year":"2004","journal-title":"Arzneimittelforschung"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1016\/j.jep.2009.11.018","article-title":"The spasmolytic activity of extracts and some isolated compounds from the leaves of Morinda morindoides (Baker) Milne-Redh. (Rubiaceae)","volume":"127","author":"Cimanga","year":"2010","journal-title":"J. Ethnopharmacol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Cerantola, S., Faggin, S., Annaloro, G., Mainente, F., Filippini, R., Savarino, E.V., Piovan, A., Zoccatelli, G., and Giron, M.C. (2021). Influence of Tilia tomentosa Moench extract on mouse small intestine neuromuscular contractility. Nutrients, 13.","DOI":"10.3390\/nu13103505"},{"key":"ref_25","first-page":"58","article-title":"Toxicological, antidiarrheal and spasmolytic activities of Solanum paniculatum","volume":"82","author":"Pereira","year":"2016","journal-title":"Planta Med."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Duangjai, A., Rawangkan, A., Yosboonruang, A., Ontawong, A., Saokaew, S., Goh, B.-H., Suganuma, M., and Phisalprapa, P. (2024). Antispasmodic activity of light-roasted coffee extract and its potential use in gastrointestinal motility disorders. Foods, 13.","DOI":"10.3390\/foods13152307"},{"key":"ref_27","first-page":"100172","article-title":"Investigation of the vasodilatory, tocolytic and spasmolytic effects of rutin on Wistar rats tissues","volume":"6","author":"Batista","year":"2025","journal-title":"Pharmacol. Res. Nat. Prod."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Kiti\u0107, N., \u017divkovi\u0107, J., \u0160avikin, K., Randjelovi\u0107, M., Jovanovi\u0107, M., Kiti\u0107, D., Miladinovi\u0107, B., Milutinovi\u0107, M., Stojiljkovi\u0107, N., and Brankovi\u0107, S. (2024). Spasmolytic activity of Gentiana lutea L. root extracts on the rat ileum: Underlying mechanisms of action. Plants, 13.","DOI":"10.3390\/plants13030453"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"205","DOI":"10.3329\/bjms.v17i2.35872","article-title":"Studies to determine antidiarrhoeal and spasmolytic activities of extract of Aegle marmelos L. fruit","volume":"17","author":"Taqvi","year":"2018","journal-title":"Bangladesh J. Med. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1104","DOI":"10.1089\/jmf.2009.0210","article-title":"Stop Fitan: Antispasmodic effect of natural extract of chestnut wood in guinea pig ileum and proximal colon smooth muscle","volume":"13","author":"Budriesi","year":"2010","journal-title":"J. Med. Food"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Zheng, Q., Chen, M., Ying, J., Wang, Z., Shan, Q., Sang, X.N., and Cao, G. (2025). The correlation between characteristics and pharmacological effects of monoterpene glycosides and tannins in Radix Paeoniae Alba. J. Pharm. Anal., 101471.","DOI":"10.1016\/j.jpha.2025.101471"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Trajkovi\u0107, M., Miladinovi\u0107, B., Mihailovi\u0107, D., Najman, S., Milutinovi\u0107, M., Ran\u0111elovi\u0107, M., Jovanovi\u0107, M., Kiti\u0107, N., \u0160avikin, K., and Kiti\u0107, D. (2026). Bioactivity of Ribes nigrum L. juice and waste extracts: Chemical composition, antioxidant, and anti-proliferative properties. Plants, 15.","DOI":"10.3390\/plants15030356"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"621","DOI":"10.1016\/j.sajb.2022.07.045","article-title":"The benefits of Salvia sclarea L. ethanolic extracts on gastrointestinal and respiratory spasms","volume":"150","year":"2022","journal-title":"S. Afr. J. Bot."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Valero, M.S., L\u00f3pez, V., Castro, M., G\u00f3mez-Rinc\u00f3n, C., Arruebo, M.P., Les, F., and Plaza, M.\u00c1. (2022). Involvement of NO\/cGMP signaling pathway, Ca2+ and K+ channels on spasmolytic effect of everlasting flower polyphenolic extract (Helichrysum stoechas (L.) Moench). Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232214422"}],"container-title":["Plants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2223-7747\/15\/3\/504\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,6]],"date-time":"2026-02-06T11:41:08Z","timestamp":1770378068000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2223-7747\/15\/3\/504"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,6]]},"references-count":34,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,2]]}},"alternative-id":["plants15030504"],"URL":"https:\/\/doi.org\/10.3390\/plants15030504","relation":{},"ISSN":["2223-7747"],"issn-type":[{"value":"2223-7747","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,6]]}}}