Inhibitory Activity on Xanthine Oxidase and Antioxidant Properties of Teucrium polium


The antioxidative activities of subfractions; methanol (CE), chloroform (CHE) and ethyl acetate (EAE) of Teucrium polium extracts (TPE) were investigated. HPLC analysis of the plant revealed the existence of procyanidins B1 and B2, gallic acid, catechin and epicatechin. All the extracts showed inhibitory properties on xanthine oxidase, with IC50 ranging from 0.80 ± 0.07 to 11.76 ± 0.50 μM/quercetin equivalent. In the cellular system, all the extracts showed a protec-tive effect greater than those of quercetin, rutin and gallic acid against t-BHP induced oxidative damages in human erythrocytes. These results were clearly confirmed by a modified thiobarbituric acid-reactive species (TBARS), and β-carotene/linoleic acid assay which demonstrated that CHE possess an inhibition ratio of the linoleic acid oxidation (83.11%) close to that of BHT (96.77%). In addition, the results showed that the extracts possess a potent DPPH radical scavenging activity and gave a reduction power greater than rutin, quercetin, gallic acid and ascorbic acid in FRAP assay. These results show that Teucrium polium extracts have strong antioxidant effects and may have some clinical benefits.

Share and Cite:

Boumerfeg, S. , Baghiani, A. , Djarmouni, M. , Ameni, D. , Adjadj, M. , Belkhiri, F. , Charef, N. , Khennouf, S. and Arrar, L. (2012) Inhibitory Activity on Xanthine Oxidase and Antioxidant Properties of Teucrium polium. Chinese Medicine, 3, 30-41. doi: 10.4236/cm.2012.31006.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] B. Halliwell and J. M. C. Gutteridge, “Free Radicals in Biology and Medicine,” 3rd Edition, Oxford University Press, London, 1999.
[2] J. L. Farber, “Mechanisms of Cell Injury by Activated Oxygen Species,” Environmental Health Perspectives, Vol. 102, 1994, pp. 17-24.
[3] B. Halliwell, “Free Radicals, Antioxidants, and Human Disease: Curiosity, Cause, or Consequence?” Lancet, Vol. 344, No. 8924, 1994, pp. 721-724. doi:10.1016/S0140-6736(94)92211-X
[4] R. Harrison, “Physiological Roles of Xanthine Oxidoreductase,” Drug Metabolism Reviews, Vol. 36, 2004, pp. 363-375. doi:10.1081/DMR-120037569
[5] H. M. Martin, J. T. Hancock, V. Salisbury and R. Harrison, “Role of Xanthine Oxidoreductase as an Antimicrobial Agent,” Infection and Immunity, Vol. 72, No. 9, 2004, pp. 4933-4939. doi:10.1128/IAI.72.9.4933-4939.2004
[6] B. T. Emmerson, “The Management of Gout,” The New England Journal of Medicine, Vol. 334, 1996, pp. 445-451. doi:10.1056/NEJM199602153340707
[7] S. L. Wallach, “The Side Effects of Allopurinol,” Hospital Practice, Vol. 33, 1998, p. 22.
[8] A. Bouloc, P. Reygagne, P. Lecoz and L. Dubertret, “Perforating Foot Ulceration with Allopurinol Therapy,” Clinical and Experimental Dermatology, Vol. 21, No. 5, 1996, pp. 351-352. doi:10.1111/j.1365-2230.1996.tb00121.x
[9] C. A. Rice-Evans, N. J. Miller and G. Paganga, “Antioxidants Properties of Phenolic Compounds,” Trends in Plant Science, Vol. 2, No. 4, 1997, pp. 152-159. doi:10.1016/S1360-1385(97)01018-2
[10] U. Plitmann, C. Heyn, A. Daniu and A. Shmida, “Pictorial Flora of Israel,” Massada, Israel, 1982.
[11] S. Facciola, “Cornucopia—A Source Book of Edible Plants,” Kampong Publications, Vista, 1990.
[12] G. Autore, F. Capasso, R. De Fusco, M. P. Fasulo, M. Lembo, N. Mascolo and A. Menghini, “Antipyretic and Antibacterial Actions of Teucrium polium,” Pharmacological Research Communications, Vol. 16, 1984, pp. 21-29. doi:10.1016/S0031-6989(84)80101-0
[13] F. Capasso, R. Cerri, P. Morrica and F. Senatore, “Chemical Composition and Anti-Inflammatory Activity of an Alcoholic Extract of Teucrium polium,” Bollettino-Societa Italiana Biologia Sperimentale, Vol. 59, No. 11, 1983, pp. 1639-1643.
[14] M. Abdollahi, H. Karimpour and H. R. Monsef-Esfehani, “Antinociceptive Effects of Teucrium polium L. Total Extract and Essential Oil in Mouse Writhing Test,” Pharmacology Research, Vol. 48, 2003, pp. 31-35.
[15] A. Alkofahi and A. H. Atta, “Pharmacological Screening of the Anti-Ulcerogenic Effects of Some Jordanian Medicinal Plants in Rats,” Journal of Ethnopharmacology, Vol. 67, No. 3, 1999, pp. 341-345. doi:10.1016/S0378-8741(98)00126-3
[16] T. Baluchnejadmojarad, M. Roghani and F. Roghani-Dehkordi, “Antinociceptive Effects of Teucrium polium Leaf Extract in the Diabetic Rat Formalin Test,” Journal of Ethnopharmacology, Vol. 97, 2005, pp. 207-210. doi:10.1016/j.jep.2004.10.030
[17] A. M. Esmaeili and R. Yazdanparast, “Hypoglycaemic Effects of Teucrium polium: Studies with Rat Pancreatic Islets,” Journal of Ethnopharmacology, Vol. 95, No. 1, 2004, pp. 27-30. doi:10.1016/j.jep.2004.06.023
[18] M. S. Suleiman, A. S. Abdul-Ghani, S. Al-Khalil and R. Amir, “Effect of Teucrium polium Boiled Leaf Extract on Intestinal Motility and Blood Pressure,” Journal of Ethnopharmacology, Vol. 22, No. 1, 1988, pp. 111-116. doi:10.1016/0378-8741(88)90236-X
[19] H. R. Rasekh, M. J. Khoshnood-Mansourkhani and M. Kamalinejad, “Hypolipidemic Effects of Teucrium polium in Rats,” Fitohterapia, Vol. 72, No. 8, 2001, pp. 937-939. doi:10.1016/S0367-326X(01)00348-3
[20] M. B. Aqel, M. N. Garaibeh and A. S. Salhab, “The Calcium Antagonistic Effect of the Volatile Oil of Teucrium polium,” International Journal of Crude Drugs Research, Vol. 28, No. 3, 1990, pp. 201-207.
[21] K. R. Markham, “Techniques of Flavonoid Identification,” London Academic Press, London, 1982.
[22] H. D. Graham, “Modified Prussian Blue Assay for Total Phenols,” Journal of Agricultural Food Chemistry, Vol. 40, No. 5, 1992, pp. 801-805. doi:10.1021/jf00017a018
[23] T. Bahorun, B. Gressier, F. Trotin, C. Brunet, T. Din, J. Vasseur, J. C. Gazin, M. Pinkas, M. Luyckx and M. Gazin, “Oxygen Species Scavenging Activity of Phenolic Extract from Howthorn Fresh Plant Organs and Pharmaceutical Preparation,” Arzneimittelforschung/Drug Research, Vol. 46, No. 11, 1996, pp. 1086-1089.
[24] Y. Li, G. Tanner and P. Larkin, “The DMACA-HCl Protocol and the Threshold Proanthocyanidin Content for Bloat Safety in Forage Legumes,” Journal of Sciences of Food Agriculture, Vol. 70, No. 1, 1996, pp. 89-101. doi:10.1002/(SICI)1097-0010(199601)70:1<89::AID-JSFA470>3.0.CO;2-N
[25] A. Scalbert, B. Monties and G. Janin, “Tannins in Wood: Comparison of Different Estimation Methods,” Journal of Agricultural Food Chemistry, Vol. 37, No. 5, 1989, pp. 1324-1329. doi:10.1021/jf00089a026
[26] J. Robak and R. J. Gryglewski, “Flavonoids Are Scavengers of Superoxide Anions,” Biochemistry and Pharmacology, Vol. 37, No. 5, 1988, pp. 837-841. doi:10.1016/0006-2952(88)90169-4
[27] M. Nishikimi, N. A. Rao and K. Yagi, “The Occurrence of Superoxide Anion in the Reaction of Reduced Phenazine Methosulfate and Molecular Oxygen,” Biochemical and Biophysical Research Communications, Vol. 46, No. 2, 1972, pp. 849-854. doi:10.1016/S0006-291X(72)80218-3
[28] V. Manna, P. Galletti, V. Cucciolla, G. Montedoro and V. Zappia, “Olive Oil Hydroxytyrosol Protects Human Erythrocytes against Oxidative Damages,” The Journal of Nutritional Biochemistry, Vol. 10, No. 3, 1999, pp. 159-165. doi:10.1016/S0955-2863(98)00085-0
[29] H. Ohkowa, N. Ohisi and K. Yagi, “Assay for Lipid Peroxides in Animal Tissue by Thiobarbituric Acid Reaction,” Analytical Biochemistry, Vol. 95, No. 2, 1979, pp. 351-358. doi:10.1016/0003-2697(79)90738-3
[30] A. Dapkevicius, R. Venskutonis, T. A. Van Beek and P. H. Linssen, “Antioxidant Activity of Extracts Obtained by Different Isolation Procedures from Some Aromatic Herbs Grown in Lithuania,” Journal of the Science of Food and Agriculture, Vol. 77, No. 1, 1998, pp. 140-146. doi:10.1002/(SICI)1097-0010(199805)77:1<140::AID-JSFA18>3.0.CO;2-K
[31] M. Burits and F. Bucar, “Antioxidant Activity of Nigella Sativa Essential Oil,” Phytotheraphy Research, Vol. 14, No. 5, 2000, pp. 323-328. doi:10.1002/1099-1573(200008)14:5<323::AID-PTR621>3.0.CO;2-Q
[32] E. A. Decker and B. Welch, “Role of Feritin as Lipid Oxidation Catalyst in Muscle Food,” Journal of Agricultural and Food Chemistry, Vol. 36, No. 3, 1990, pp. 674-677. doi:10.1021/jf00093a019
[33] R. Pulido, L. Bravo and F. Saura-Calixto, “Antioxidant Activity of Dietary Polyphenols as Determined Modified Ferric Reducing/Antioxidant Power Assay,” Journal of Agricultural Food Chemistry, Vol. 48, No. 8, 2000, pp. 3396-3402. doi:10.1021/jf9913458
[34] J. M. McCord and I. Fridovich, “The Reduction of Cytochrome C by Milk Xanthine Oxidase,” Journal of Biological Chemistry, Vol. 243, 1968, pp. 5753-5760.
[35] P. G. Avis, F. Bergel, R. C. Bray, D. W. F. James and K. V. Shooter, “Cellular Constituent, the Chemistry of Xanthine Oxidase. Part II: The Homogeneity of Crystalline Metalloflavoproteine Fraction,” Journal of Chemical Society, 1956, pp. 1212-1219. doi:10.1039/jr9560001212
[36] S. Boumerfeg, A. Baghiani, D. Messaoudi, S. Khennouf and L. Arrar, “Antioxidant Properties and Xanthine Oxidase Inhibitory Effects of Tamus communis L. Root Extracts,” Phytotherapy Research, Vol. 23, No. 2, 2009, pp. 283-288. doi:10.1002/ptr.2621
[37] A. Baghiani, S. Boumerfeg, F. Belkhiri, S. Khennouf, N. Charef, D. Harzallah, L. Arrar and M. Abdel-Wahhab, “Antioxidant and Radical Scavenging Properties of Carthamus caeruleus L. Extracts Grow Wild in the Algeria Flora,” Comunicata Scientiae, Vol. 1, No. 2, 2010, pp. 128-136.
[38] D. O. Kim, K. W. Lee, H. J. Lee and C. Y. Lee, “Vitamin C Equivalent Antioxidant Capacity (VCEAC) of Phenolic Phytochemicals,” Journal of Agricultural and Food Chemistry, Vol. 50, No. 13, 2002, pp. 3713-3717. doi:10.1021/jf020071c
[39] F. N. Ko, G. Hsiao and Y. H. Kuo, “Protection of Oxidative Hemolysis by Demethydiisoeugenol in Normal and Beta-Thalassemic Red Blood Cells,” Free Radical Biology and Medicine, Vol. 22, No. 1-2, 1997, pp. 215-222. doi:10.1016/S0891-5849(96)00295-X
[40] H. Kitagawa, H. Sakamoto and Y. Tano, “Inhibitory Effects of Flavonoids on Free Radical-Induced Hemolysis and Their Oxidative Effects on Hemoglobin,” Chemical and Pharmaceutical Bulletin, Vol. 52, 2004, pp. 999-1001. doi:10.1248/cpb.52.999
[41] J. Zhishen, T. Mengcheng and W. Jianming, “The Determination of Flavonoid Contents in Mulberry and Their Scavenging Effects on Superoxide Radicals,” Food Chemistry, Vol. 64, No. 4, 1999, pp. 555-559. doi:10.1016/S0308-8146(98)00102-2
[42] S. B. Lotitio and C. G. Fraga, “Catechin Prevents Human Plasma Oxidation,” Free Radical Biology and Medicine, Vol. 24, 1998, pp. 435-441. doi:10.1016/S0891-5849(97)00276-1
[43] S. M. Suboh, Y. Y. Bilto, T. A. Aburjai, “Protective Effects of Selected Medicinal Plants against Protein Degradation, Lipid Peroxidation and Deformability Loss of Oxidatively Stressed Human Erythrocytes,” Phytotherapy Research, Vol. 18, No. 4, 2004, pp. 280-284. doi:10.1002/ptr.1380
[44] P. Ljubuncic, H. Azaizeh, I. Portnaya, U. Cogan, O. Said, K. A. Saleh, K. Abu Saleh and A. Bomzon, “Antioxidant Activity and Cytotoxicity of Eight Plants Used in Traditional Arab Medicine in Israel,” Journal of Ethnopharmacology, Vol. 99, No. 1, 2005, pp. 43-47. doi:10.1016/j.jep.2005.01.060
[45] Y. H. Chu, C. L. Chang and H. F. Hsu, “Flavonoid Content of Several Vegetables and Their Antioxidant Activity,” Journal of the Science of Food and Agriculture, Vol. 80, No. 5, 2000, pp. 561-566. doi:10.1002/(SICI)1097-0010(200004)80:5<561::AID-JSFA574>3.0.CO;2-#
[46] A. M. Nuutila, R. Puupponen-Pimia, M. Aarni and K. M. Oksman-Caldentey, “Comparison of Antioxidant Activities of Onion and Garlic Extracts by Inhibition of Lipid Peroxidation and Radical Scavenging Activity,” Food Chemistry, Vol. 81, No. 4, 2003, pp. 485-493. doi:10.1016/S0308-8146(02)00476-4
[47] G. A. Agbor, J. E. Oben, J. Y. Ngogang, C. Xin and J. A. Vinson, “Antioxidant Capacity of Some Herbs/Spices from Cameroon: A Comparative Study of Two Methods,” Journal of Agricultural and Food Chemistry, Vol. 53, No. 17, 2005, pp. 6819-6824. doi:10.1021/jf050445c
[48] S. Demiray, M. E. Pintado and P. M. L. Castro, “Evaluation of Phenolic Profiles and Antioxidant Activities of Turkish Medicinal Plants: Tilia Argentea, Crataegi Folium Leaves and Polygonum Bistorta Roots,” World Academy of Science, Engineering and Technology, Vol. 54, 2009, pp. 312-317.
[49] C. A. Hall and S. L. Cuppett, “Activities of Natural Antioxidants,” In: O. I. Aruoma and S. L. Cuppett, Eds., Antioxidant Methodology in Vivo and in Vitro Concepts, AOCS Press, Champaign, 1997, pp. 2-29.
[50] D. Ferreira, D. Slade and J. P. J. Marais, “Flavans and Proanthocyanidins,” In: O. M. Anderson and K. R. Markham, Eds., Flavonoids: Chemistry, Biochemistry and Applications, CRC Press, Boca Raton, 2006, pp. 553-616.
[51] E. N. Frankel and A. S. Meyer, “The Problems of Using One-Dimensional Methods to Evaluate Multifunctional Food and Biological Antioxidants,” Journal of the Science of Food and Agriculture, Vol. 80, No. 13, 2000, pp. 1925-1940. doi:10.1002/1097-0010(200010)80:13<1925::AID-JSFA714>3.0.CO;2-4
[52] C. M. Liyana-Pathirana and F. Shahidi, “Antioxydant Propreties of Commercial Soft and Hard Winter Wheats (Triticum aestivium L.) and Their Milling Fractions,” Journal of the Science of Food and Agriculture, Vol. 86, No. 3, 2006, pp. 477-485. doi:10.1002/jsfa.2374
[53] T. Kadifkova-Panovska, S. Kulevanova and M. Stefova, “In Vitro Antioxidant Activity of Some Teucrium Species (Lamiaceae),” Acta Pharmaceutica, Vol. 55, 2005, pp. 207-214.
[54] C. Sánchez-Moreno, “Methods Used to Evaluate the Free Radical Scavenging Activity in Foods and Biological Systems,” Food Science and Technology International, Vol. 8, 2002, pp. 121-137.
[55] V. Bondent, W. Brand-Williams and C. Bereset, “Kinetic and Mechanism of Antioxidant Activity Using the DPPH Free Radical Methods,” Lebensmittel Wissenschaft and Technologie, Vol. 30, No. 6, 1997, pp. 609-615. doi:10.1006/fstl.1997.0240
[56] C. A. Rice-Evans, N. J. Miller and G. Paganga, “Structure, Antioxidant Activity Relationship of Flavonoids and Phenolic Acids,” Free Radical Biology and Medicine, Vol. 20, No. 7, 1996, pp. 933-956. doi:10.1016/0891-5849(95)02227-9
[57] I. F. Benzie and J. J. Strain, “The Ferric Reducing Ability of Plasma (FRAP) as a Measure of ‘Antioxidant Power’: The FRAP Assay,” Analytical and Bioanalytical Chemistry, Vol. 239, 1996, pp. 70-76.
[58] Z. Maksimovi?, D. Malen?i? and N. Kova?evi?, “Polyphenol Contents and Antioxidant Activity of Maydis Stigma Extracts,” Bioresource Technology, Vol. 96, No. 8, 2005, pp. 873-877. doi:10.1016/j.biortech.2004.09.006
[59] W. Wang and M. T. Goodman, “Antioxidant Property of Dietary Phenolic Agents in a Human LDL-Oxidation ex Vivo Model: Interaction of Protein Binding Activity,” Nutrition Research, Vol. 19, No. 2, 1999, pp. 191-202. doi:10.1016/S0271-5317(98)00183-3
[60] H. Azaizeh, P. Ljubuncic, I. Portnaya, O. Said, U. Cogan and A. Bomzon, “Fertilization-Induced Changes in Growth Parameters and Antioxidant Activity of Medicinal Plants Used in Traditional Arab Medicine,” Evidence-Based Complementary and Alternative Medicine, Vol. 2, 2005, pp. 549-556. doi:10.1093/ecam/neh131
[61] A. Ardestani and R. Yazdanparast, “Inhibitory Effects of Ethyl Acetate Extract of Teucrium polium on in Vitro Protein Glycoxidation,” Food and Chemical Toxicology, Vol. 45, No. 12, 2007, pp. 2402-2411. doi:10.1016/j.fct.2007.06.020
[62] D. Bagchi, A. Garg, R. L. Krohn , M. Bagchi, J. Balmoori and S. J. Stohs, “Protective Effects of Grape Seed Proanthocyanidins and Selected Antioxidants against TPA-Induced Hepatic and Brain Lipid Peroxidation and DNA Fragmentation, and Peritoneal Macrophage Activation in Mice,” General Pharmacology, Vol. 30, No. 6, 1998, pp. 771-776. doi:10.1016/S0306-3623(97)00332-7
[63] M. Murray and J. Pizzorno, “Procyanidolic Oligomers,” In: M. Murray and J. Pizzorno, Eds., The Textbook of Natural Medicine, 2nd Edition, Churchill Livingston, London, 1999, pp. 899-902.
[64] D. Bagchi, R. L. Krohn, M. Bagchi, M. X. Tran and S. J. Stohs, “Oxygen Free Radical Scavenging Abilities of Vitamins C and E, and a Grape Seed Proanthocyanidin Extract in Vitro,” Research Communications in Molecular Pathology & Pharmacology, Vol. 95, 1997, pp. 179-189.
[65] E. Bombardelli, P. Morazzoni, M. Carini, G. Aldini and F. R. Maffei, “Biological Activity of Procyanidins from Vitis vinifera L.,” BioFactors, Vol. 6, No. 4, 1997, pp. 429-431. doi:10.1002/biof.5520060411
[66] S. Son and B. A. Lewis, “Free Radical Scavenging and Antioxidant Activity of Caffeic Acid Amide and Ester Analogues: Structure-Activity Relationship,” Journal of Agricultural and Food Chemistry, Vol. 50, No. 3, 2002, pp. 468-472. doi:10.1021/jf010830b
[67] W. Zheng and S. Y. Wang, “Antioxidant Activity and Phenolic Compounds in Selected Herbs,” Journal of Agricultural and Food Chemistry, Vol. 49, No. 11, 2001, pp. 5165-5170. doi:10.1021/jf010697n

Copyright © 2024 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.