Radioprotective Properties of the Phytochemically Characterized Extracts of Crataegus monogyna, Cornus mas and Gentianella austriaca on Human Lymphocytes in vitro

Abstract

The objective of the present study is to evaluate the radioprotective properties of extracts of Crataegus monogyna Jacq. (Rosaceae) fruit, Cornus mas L. (Cornaceae) leaves and Gentianella austriaca (A. Kern. & Jos. Kern.) Holub (Gentianaceae) aerial parts on cultured human peripheral blood lymphocytes in vitro after irradiation with 2 Gy of ⁶⁰Co γ-rays. Plants were collected at Mt. Maljen in Serbia, air-dried and powdered, and the total phenolic content was analyzed. In C. mas leaves, ellagic and gallic acid were found to be the dominant compounds, whereas C. monogyna fruit was rich in procyanidins and flavonoids. The main constituents of G. austriaca aerial parts were γ-pyrones and secoiridoids. Cell cultures were treated with four different doses of plant extracts (0.1 mg/mL to 0.4 mg/mL). Treatment with the lowest dose gave the best protective effect. Significant radiorecovery potentials of C. mas and C. monogyna were observed, seen as a reduced incidence of radiation-induced micronuclei (19.23 % and 13.24 %, respectively), reduced levels of lipid peroxidation products (50.04 % and 13.18 %, respectively) and two-fold enhanced apoptosis. Both extracts slowed down cell proliferation gradually, enabling more time for repair. G. austriaca possesses strong antioxidant properties, significantly reducing lipid peroxidation and the incidence of micronuclei (for 30.88 % and 35.56 %, respectively) while enhancing apoptosis with no perturbation of the cell cycle. This study may contribute to the search for novel radioprotective agents.

Key words

Crataegus monogyna - Rosaceae - Cornus mas - Cornaceae - Gentianella austriaca - Gentianaceae - polyphenols - radioprotection

References

• 1 Cos P, Calomme M, Sindambiwe J B, De Bruyne T, Cimanga K, Pieters L. et al . Cytotoxicity and lipid peroxidation-inhibiting activity of flavonoids.  Planta Med. 2001;  67 515-9.
• 2 Nijveldt R J, van Nood E, van Hoorn D EC, Boelens P G, van Norren K, van Leeuwen P AM. Flavonoids: a review of probable mechanisms of action and potential applications.  Am J Clin Nutr. 2001;  74 418-25.
• 3 Schuchter L M, Glik J. The current status of WR-2721 (amifostine): a chemotherapy and radiation therapy protector.  Biol Ther Cancer. 1993;  3 1-10.
• 4 Sagar R K, Chawla R, Arora R, Singh S, Krishna B, Sharma R K. et al . Protection of the hemopoietic system by Podophyllum hexandrum against gamma radiation-induced damage.  Planta Med. 2006;  72 114-20.
• 5 Arora R, Gupta D, Chawla R, Sagar R, Sharma A, Kumar R. et al . Radioprotection by plant products: present status and future prospects.  Phytother Res. 2005;  19 1-22.
• 6 Nadinic E, Gorzalczany S, Rojo A, van Baren C, Debenedetti S, Acevedo C. Topical anti-inflammatory activity of Gentianella achalensis .  Fitoterapia. 1999;  70 166-71.
• 7 Natarajan A T, Boei J J, Darroudi F, Van Diemen P C, Dulout F, Hande M P. et al . Current cytogenetic methods for detecting exposure and effects of mutagens and carcinogens.  Environ Health Perspect. 1996;  104 445-8.
• 8 West C M. Invited review: intrinsic radiosensitivity as a predictor of patient response to radiotherapy.  Br J Radiol. 1995;  68 827-37.
• 9 Singleton V L, Rossi J A. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents.  Am J Enol Vitic. 1965;  16 144-58.
• 10 European Pharmacopoeia. 5th Edition, Vol. 2 Strasbourg; Council of Europe 2005: 1712-3.
• 11 Jankovic T, Krstic D, Aljancic I, Savikin-Fodulovic K, Menkovic N, Vajs V. et al . Xanthones and C-glucosides from the aerial parts of four species of Gentianella from Serbia and Montenegro.  Biochem Syst Ecol. 2005;  33 729-35.
• 12 Law on health care. Official Gazette of the Republic of Serbia. Vol. 107 Parliament of the Republic of Serbia 2005: 112-61.
• 13 International Atomic Energy Agency: Cytogenetic analysis for Radiation Dose Assessment. Technical Series No. 405. Vienna; IAEA 2001.
• 14 Fenech M. The cytokinesis-block micronucleus technique: a detailed description of the method and its application to genotoxicity studies in human populations.  Mutat Res. 1993;  285 35-44.
• 15 Surralles J, Xamena N, Creus A, Marcos R. The suitability of the micronucleus assay in human lymphocytes as a new biomarker of excision repair.  Mutat Res. 1995;  342 43-59.
• 16 Aruoma O I, Halliwell B, Laughton M J, Quinlan G J, Gutteridge J M. The mechanism of initiation of lipid peroxidation. Evidence against a requirement for an iron(II)-iron(III) complex.  Biochem J. 1989;  258 617-20.
• 17 Lowry O H, Rosebrough N J, Farr A L, Randall R J. Protein measurement with the Folin phenol reagent.  J Biol Chem. 1951;  193 265-75.
• 18 Crompton N E, Ozsahin M. A versatile and rapid radiosensitivity assay of peripheral blood leukocytes based on DNA and surface-marker assessment of cytotoxicity.  Radiat Res. 1997;  147 55-60.
• 19 Thresiamma K C, George J, Kuttan R. Protective effect of curcumin, ellagic acid and bixin on radiation induced genotoxicity.  J Exp Clin Cancer Res. 1998;  17 431-4.
• 20 Gebhardt R. Oxidative stress, plant-derived antioxidants and liver fibrosis.  Planta Med. 2002;  68 289-96.
• 21 Laupattarakasem P, Houghton P J, Hoult J RS. Anti-inflammatory isoflavonoids from the stems of Derris scandens .  Planta Med. 2004;  70 496-501.
• 22 Seghrouchni I, Drai J, Bannier E, Riviere J, Calmard P, Garcia I. et al . Oxidative stress parameters in type I, type II and insulin-treated type 2 diabetes mellitus; insulin treatment efficiency.  Clin Chim Acta. 2002;  321 89-96.
• 23 Das U. A radical approach to cancer.  Med Sci Monit. 2002;  8 79-92.
• 24 Umegaki K, Ikegami S, Inoue K, Ichikawa T, Kobayashi G, Soeno N. et al . Beta carotene prevents X-ray induction of micronuclei in human lymphocytes.  Am J Clin Nutr. 1994;  59 409-12.
• 25 Vrinda B, Uma Devi P. Radiation protection of human lymphocyte chromosomes in vitro by orientin and vicenin.  Mutat Res. 2001;  498 39-46.
• 26 Jagetia G C, Venkatesh P, Baliga M S. Evaluation of the radioprotective effect of Aegle marmelos (L.) Correa in cultured human peripheral blood lymphocytes exposed to different doses of gamma-radiation: a micronucleus study.  Mutagenesis. 2003;  18 387-93.
• 27 Wen X, Walle T. Methylated flavonoids have greatly improved intestinal absorption and metabolic stability.  Drug Metab Dispos. 2006;  34 786-92.