{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T17:14:31Z","timestamp":1792602871641,"version":"4.5.0"},"reference-count":49,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2023,6,25]],"date-time":"2023-06-25T00:00:00Z","timestamp":1687651200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"European Union-NextGenerationEU","award":["BG-RRP-2.004-0004-C01"],"award-info":[{"award-number":["BG-RRP-2.004-0004-C01"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>An in vitro\/in vivo hepatotoxicity and hepatoprotection evaluation of a defatted extract and a phenolic fraction from Phlomis tuberosa, administered alone and in a carbon tetrachloride (CCl4)-induced metabolic bioactivation model, was performed. The extract and the phenolic fraction were analysed by high performance liquid chromatography (HPLC) to determine the total flavonoid content, to identify flavonoids and to quantify verbascoside. In addition, total polyphenolics in the samples were expressed as gallic acid equivalents. Applied alone, the extract and the fraction (5, 10 and 50 \u00b5g\/mL) did not show a statistically significant hepatotoxic effect on isolated rat hepatocytes in vitro. In a CCl4-induced hepatotoxicity model, the samples exhibited a concentration-dependent, statistically significant hepatoprotective effect, which was most pronounced at 50 \u00b5g\/mL for both. The phenolic fraction exhibited a more pronounced hepatoprotective effect compared to the extract. Data from the in vitro study on the effects of the extract were also confirmed in the in vivo experiment conducted in a CCl4-induced hepatotoxicity model in rats. A histopathological study showed that the animals treated with CCl4 and the extract had an unaltered histoarchitecture of the liver. The effects of the extract were the same as those of silymarin.<\/jats:p>","DOI":"10.3390\/ijms241310631","type":"journal-article","created":{"date-parts":[[2023,6,26]],"date-time":"2023-06-26T01:42:32Z","timestamp":1687743752000},"page":"10631","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["In Vitro\/In Vivo Hepatoprotective and Antioxidant Effects of Defatted Extract and a Phenolic Fraction Obtained from Phlomis Tuberosa"],"prefix":"10.3390","volume":"24","author":[{"given":"Magdalena","family":"Kondeva-Burdina","sequence":"first","affiliation":[{"name":"Laboratory of Drug Metabolism and Drug Toxicity, Department of Pharmacology, Pharmacotherapy and Toxicology, Faculty of Pharmacy, Medical University of Sofia, 2 Dunav st., 1000 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5091-6058","authenticated-orcid":false,"given":"Aleksandar","family":"Shkondrov","sequence":"additional","affiliation":[{"name":"Department of Pharmacognosy, Faculty of Pharmacy, Medical University of Sofia, 2 Dunav st., 1000 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Georgi","family":"Popov","sequence":"additional","affiliation":[{"name":"Department of Non-infectious Diseases, Pathology and Pharmacology, Faculty of Veterinary Medicine, University of Forestry, 1000 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vasil","family":"Manov","sequence":"additional","affiliation":[{"name":"Department of Non-infectious Diseases, Pathology and Pharmacology, Faculty of Veterinary Medicine, University of Forestry, 1000 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8559-0369","authenticated-orcid":false,"given":"Ilina","family":"Krasteva","sequence":"additional","affiliation":[{"name":"Department of Pharmacognosy, Faculty of Pharmacy, Medical University of Sofia, 2 Dunav st., 1000 Sofia, Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1582","DOI":"10.1016\/j.biotechadv.2015.08.001","article-title":"Discovery and Resupply of Pharmacologically Active Plant-Derived Natural Products: A Review","volume":"33","author":"Atanasov","year":"2015","journal-title":"Biotechnol. Adv."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/j.jep.2009.06.022","article-title":"Phytochemistry and Biological Activities of Phlomis Species","volume":"125","author":"Amor","year":"2009","journal-title":"J. Ethnopharmacol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1002\/ptr.1261","article-title":"Screening of Some Greek Aromatic Plants for Antioxidant Activity","volume":"17","author":"Couladis","year":"2003","journal-title":"Phytother. Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1007\/s11418-008-0255-y","article-title":"Free-Radical-Scavenging Principles from Phlomis Caucasica","volume":"62","author":"Delazar","year":"2008","journal-title":"J. Nat. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1002\/ffj.1427","article-title":"Composition of the Volatile Oils of Phlomis Lanceolata Boiss. & Hohen., Phlomis Anisodonta Boiss. and Phlomis Bruguieri Desf. from Iran","volume":"20","author":"Sarkhail","year":"2005","journal-title":"Flavour Fragr. J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1155\/2014\/706302","article-title":"Some in Vitro\/in Vivo Chemically-Induced Experimental Models of Liver Oxidative Stress in Rats","volume":"2014","author":"Simeonova","year":"2014","journal-title":"Biomed. Res. Int."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1016\/S0165-1110(96)00027-9","article-title":"Carbon Tetrachloride: Genetic Effects and Other Modes of Action","volume":"366","author":"McGregor","year":"1996","journal-title":"Mutat. Res. Genet. Toxicol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"61","DOI":"10.3897\/pharmacia.70.e97050","article-title":"Essential Oil in Phlomis Tuberosa","volume":"70","author":"Shkondrov","year":"2023","journal-title":"Pharmacia"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1007\/s10600-010-9602-9","article-title":"Chemical Composition of Essential Oils from Galeopsis Bifida and Phlomoides Tuberosa","volume":"46","author":"Olennikov","year":"2010","journal-title":"Chem. Nat. Compd."},{"key":"ref_10","first-page":"363","article-title":"Determination of Flavonoids in Related Astragalus Species (Sect. Incani) Occurring in Bulgaria","volume":"70","author":"Shkondrov","year":"2017","journal-title":"Comptes. Rendus. L\u2019Academie Bulg. Des Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/S0165-9936(04)00201-8","article-title":"Validation of Qualitative Analytical Methods","volume":"23","author":"Trullols","year":"2004","journal-title":"TrAC Trends Anal. Chem."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Chan, C.C., and Saraswat, P. (2010). Analytical Method Verification, Method Revalidation, and Method Transfer, John Wiley & Sons, Inc.","DOI":"10.1002\/9780470630716.ch3"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Andersen, O.M., and Markham, K.R. (2005). Flavonoids: Chemistry, Biochemistry and Applications, CRC Press.","DOI":"10.1201\/9781420039443"},{"key":"ref_14","unstructured":"Guideline, I.C.H.H.T. (, January November). Validation of Analytical Procedures: Text and Methodology Q2 (R1). Proceedings of the International conference on harmonization, Geneva, Switzerland."},{"key":"ref_15","first-page":"360","article-title":"Spectrophotometric Determination of the Total Phenolic Content, Spectral and Fluorescence Study of the Herbal Unani Drug Gul-e-Zoofa (Nepeta Bracteata Benth)","volume":"12","author":"Siddiqui","year":"2017","journal-title":"J. Taibah Univ. Med. Sci."},{"key":"ref_16","first-page":"69","article-title":"Mechanism for the Hepatotoxicity of the Antiandrogen, Nilutamide. Evidence Suggesting That Redox Cycling of This Nitroaromatic Drug Leads to Oxidative Stress in Isolated Hepatocytes","volume":"263","author":"Fau","year":"1992","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_17","unstructured":"Timbrell, J. (2000). Principles of Biochemical Toxicology, Taylor and Francis. [3rd ed.]."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1080\/713611034","article-title":"Hepatotoxicity and Mechanism of Action of Haloalkanes: Carbon Tetrachloride as a Toxicological Model","volume":"33","author":"Weber","year":"2003","journal-title":"Crit. Rev. Toxicol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1177\/030089168907500410","article-title":"Lipid Peroxidation and Cancer: A Critical Reconsideration","volume":"75","author":"Dianzani","year":"1989","journal-title":"Tumori J."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1124\/dmd.32.6.587","article-title":"Silybin Inactivates Cytochromes P450 3A4 and 2C9 and Inhibits Major Hepatic Glucuronosyltransferases","volume":"32","author":"Sridar","year":"2004","journal-title":"Drug Metab. Dispos."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1002\/jat.2550100408","article-title":"Prevention by Silymarin of Membrane Alterations in Acute CCl4 Liver Damage","volume":"10","author":"Muriel","year":"1990","journal-title":"J. Appl. Toxicol."},{"key":"ref_22","first-page":"105","article-title":"Biochemical Bases of the Pharmacological Action of the Flavonoid Silymarin and of Its Structural Isomer Silibinin","volume":"27","author":"Valenzuela","year":"1994","journal-title":"Biol. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/j.cbi.2005.08.005","article-title":"Silibinin Inhibits Cell Invasion through Inactivation of Both PI3K-Akt and MAPK Signaling Pathways","volume":"156","author":"Chen","year":"2005","journal-title":"Chem. Biol. Interact."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2605","DOI":"10.1002\/med.21717","article-title":"Therapeutic Potential of Phenylethanoid Glycosides: A Systematic Review","volume":"40","author":"Wu","year":"2020","journal-title":"Med. Res. Rev."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"46","DOI":"10.4314\/ajtcam.v11i3.8","article-title":"Antioxidant Activity in Vitro and Hepatoprotective Effect of Phlomis Maximowiczii in Vivo","volume":"11","author":"Gu","year":"2014","journal-title":"Afr. J. Tradit. Complement. Altern. Med."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Rasheed, M.U., Naqvi, S.A.R., Rasool, N., Shah, S.A.A., and Zakaria, Z.A. (2022). Anti-Diabetic and Cytotoxic Evaluation of Phlomis Stewartii Plant Phytochemicals on Cigarette Smoke Inhalation and Alloxan-Induced Diabetes in Wistar Rats. Metabolites, 12.","DOI":"10.3390\/metabo12111133"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"219","DOI":"10.4103\/0973-1296.66940","article-title":"Effect of Phlomis Persica on Glucose Levels and Hepatic Enzymatic Antioxidants in Streptozotocin-Induced Diabetic Rats","volume":"6","author":"Sarkhail","year":"2010","journal-title":"Pharmacogn. Mag."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1016\/j.phrs.2007.07.003","article-title":"Antidiabetic Effect of Phlomis Anisodonta: Effects on Hepatic Cells Lipid Peroxidation and Antioxidant Enzymes in Experimental Diabetes","volume":"56","author":"Sarkhail","year":"2007","journal-title":"Pharmacol. Res."},{"key":"ref_29","first-page":"387","article-title":"Phlomis Fruticosa L. Exerts in Vitro Antineurodegenerative and Antioxidant Activities and Induces Prooxidant Effect in Glioblastoma Cell Line","volume":"21","author":"Stojkovic","year":"2022","journal-title":"EXCLI J."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"113884","DOI":"10.1016\/j.jpba.2020.113884","article-title":"Chemical Profiling, Antimicrobial, Anti-Enzymatic, and Cytotoxic Properties of Phlomis Fruticosa L","volume":"195","author":"Zengin","year":"2021","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2387","DOI":"10.1002\/ptr.6429","article-title":"Protective Effects Induced by Alcoholic Phlomis Fruticosa and Phlomis Herba-Venti Extracts in Isolated Rat Colon: Focus on Antioxidant, Anti-Inflammatory, and Antimicrobial Activities in Vitro","volume":"33","author":"Ferrante","year":"2019","journal-title":"Phytother. Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"e20180404","DOI":"10.1590\/0001-3765201920180404","article-title":"Evaluating Antimicrobial and Antioxidant Capacity of Endemic Phlomis Russeliana from Turkey and Its Antiproliferative Effect on Human Caco-2 Cell Lines","volume":"91","author":"Alpay","year":"2019","journal-title":"An. Acad. Bras. Cienc."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2449","DOI":"10.1080\/14786419.2019.1677650","article-title":"Iridoid Glucosides and Triterpene Acids from Phlomis Linearifolia, Growing in Uzbekistan and Its Hepatoprotective Activity","volume":"35","author":"Usmanov","year":"2021","journal-title":"Nat. Prod. Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"851","DOI":"10.1080\/14786419.2018.1502767","article-title":"Chemical Composition and Antioxidant Activity of Phlomis Leucophracta, an Endemic Species from Turkey","volume":"34","author":"Sarikurkcu","year":"2020","journal-title":"Nat. Prod. Res."},{"key":"ref_35","first-page":"282","article-title":"Chemical Composition, Antioxidant, Genotoxique and Antigenotoxic Potentials of Phlomis Bovei De No\u00e9 Aerial Parts","volume":"19","author":"Zaabat","year":"2020","journal-title":"Iran. J. Pharm. Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1080\/01480540902882200","article-title":"Screening of Antimutagenicity via Antioxidant Activity in Different Extracts from the Flowers of Phlomis Crinita Cav. Ssp Mauritanica Munby from the Center of Tunisia","volume":"32","author":"Dellai","year":"2009","journal-title":"Drug Chem. Toxicol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1016\/j.phytochem.2013.09.015","article-title":"Isolation of Natural Compounds from Phlomis Stewartii Showing \u03b1-Glucosidase Inhibitory Activity","volume":"96","author":"Jabeen","year":"2013","journal-title":"Phytochemistry"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.molimm.2019.02.002","article-title":"Phenylethanoid Glycosides of Phlomis Younghusbandii Mukerjee Ameliorate Acute Hypobaric Hypoxia-Induced Brain Impairment in Rats","volume":"108","author":"Luan","year":"2019","journal-title":"Mol. Immunol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1089\/jmf.2008.0299","article-title":"Phlomis Mauritanica Extracts Reduce the Xanthine Oxidase Activity, Scavenge the Superoxide Anions, and Inhibit the Aflatoxin B1-, Sodium Azide-, and 4-Nitrophenyldiamine-Induced Mutagenicity in Bacteria","volume":"13","author":"Limem","year":"2010","journal-title":"J. Med. Food"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1016\/j.phytochem.2015.03.007","article-title":"Phenylpropanoids and Flavonoids from Phlomis Kurdica as Inhibitors of Human Lactate Dehydrogenase","volume":"116","author":"Bader","year":"2015","journal-title":"Phytochemistry"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1007\/s11130-010-0169-1","article-title":"Antioxidant Activity and Phenylpropanoids of Phlomis lychnitis L.: A Traditional Herbal Tea","volume":"65","author":"Akerreta","year":"2010","journal-title":"Plant Foods Hum. Nutr."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1016\/j.crvi.2009.05.006","article-title":"Phenolic Composition and Antioxidant Activities of Two Phlomis Species: A Correlation Study","volume":"332","author":"Zhang","year":"2009","journal-title":"Comptes Rendus Biol."},{"key":"ref_43","unstructured":"Markham, K.R. (1982). Techniques of Flavonoid Identification, Academic Press London."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Swartz, M.E., and Krull, I.S. (2018). Analytical Method Development and Validation, CRC Press.","DOI":"10.1201\/9781315275161"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1179\/135100006X100968","article-title":"Effect of Benzophenones from Hypericum Annulatum on Carbon Tetrachloride-Induced Toxicity in Freshly Isolated Rat Hepatocytes","volume":"11","author":"Mitcheva","year":"2006","journal-title":"Redox Rep."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"17854","DOI":"10.1038\/s41598-022-20412-3","article-title":"Ameliorative Effect of Oregano (Origanum lulgare) versus Silymarin in Experimentally Induced Hepatic Encephalopathy","volume":"12","author":"Abdelghffar","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1002\/ptr.2146","article-title":"Ameliorative Effects of Pycnogenol\u00ae on Carbon Tetrachloride-Induced Hepatic Oxidative Damage in Rats","volume":"21","author":"Ahn","year":"2007","journal-title":"Phyther. Res."},{"key":"ref_48","first-page":"169","article-title":"Antioxidant and Protective Effect of an Oleanolic Acid-Enriched Extract of A. Deliciosa Root on Carbon Tetrachloride Induced Rat Liver Injury","volume":"16","author":"Bai","year":"2007","journal-title":"Asia Pac. J. Clin. Nutr."},{"key":"ref_49","unstructured":"Bancroft, J.D., and Gamble, M. (2002). Theory and Practice of Histological Techniques, Churchill Livingstone Publications. [5th ed.]."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/13\/10631\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:00:35Z","timestamp":1760126435000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/13\/10631"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,6,25]]},"references-count":49,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2023,7]]}},"alternative-id":["ijms241310631"],"URL":"https:\/\/doi.org\/10.3390\/ijms241310631","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,6,25]]}}}