Genotoxicity evaluation of hydroalcoholic and aqueous extracts of Dorema aucheri by the comet assay

Document Type : Original Article

Authors

1 Department of Pharmacology, Isfahan Pharmaceutical Sciences Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran

2 Department of Pharmacognosy, Isfahan Pharmaceutical Sciences Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: Dorema aucheri is a plant of Apiaceae family which is used widely in some states of Iran. Different extracts and essential oil of Dorema species contain flavonoids and cumarin compounds which have anti-hypertensive, cholesterol- and triglycerides-lowering properties. This study was undertaken to evaluate the genotoxic properties of hydroalcoholic and aqueous extracts of D. aucheri on human hepatoma cells using the comet assay method for safety evaluation.
Materials and Methods: In this method, after incubation of cells with different concentrations of extracts, cell suspensions were added to pre-coated normal agarose slides. After lysis, electrophoresis and neutralization process, staining was done by ethidium bromide and comets were observed using a fluorescence microscope. Tail length, percentage of DNA in tail and tail moment parameters were measured.
Results: Statistical analysis of the results demonstrated that concentrations more than 500 μg/ml of hydroalcoholic and aqueous extract of D. aucheri were genotoxic.
Conclusion: It can be concluded from the results that taking the concentrations less than these dosages of extracts are safe but more studies are required to determine genotoxic mechanisms of this plant.

Keywords

1.
Pakseresht M, Forman D, Malekzadeh R, Yazdanbod A, West RM, Greenwood DC, et al. Dietary habits and gastric cancer risk in north-west Iran. Cancer Causes Control 2011;22:725-36.  Back to cited text no. 1
    
2.
Semnani S, Sadjadi A, Fahimi S, Nouraie M, Naeimi M, Kabir J, et al. Declining incidence of esophageal cancer in the Turkmen Plain, eastern part of the Caspian Littoral of Iran: A retrospective cancer surveillance. Cancer Detect Prev 2006;30:14-9.  Back to cited text no. 2
    
3.
Wollenweber E, Dörr M, Rustiyan A. Dorema aucheri, the first umbelliferous plant found to produce exudate flavonoids. Phytochemistry 1995;38:1417.  Back to cited text no. 3
    
4.
Narasimhacharya AV, Nitesh K, Desai TN. Prosopis juliflora as an antihypercholesterolemic agent. J Herb Med Toxicol 2010;4:31-4.  Back to cited text no. 4
    
5.
Azarneushan F, Karami M, Golizadeh L, Davary K. The effect of Dorema aucheri-Hydroalcoholic extracts on thyroids hormones in adult male rats. J Shahrekord Univ Med Sci 2010;12:76-83.  Back to cited text no. 5
    
6.
Azarnioushan F, Khatamsaz S, Sadeghi HE. The effects of hydroalcoholic extract of Dorema Aucheri on blood concentration of gonadotropin and androgen hormones in adult male rats. Armaghan Danesh 2009;14:63-70.  Back to cited text no. 6
    
7.
Mozaffarieh M, Schoetzau A, Sauter M, Grieshaber M, Orgül S, Golubnitschaja O, et al. Comet assay analysis of single-stranded DNA breaks in circulating leukocytes of glaucoma patients. Mol Vis 2008;14:1584-8.  Back to cited text no. 7
    
8.
Jena GB, Kaul CL, Ramarao P. Genotoxicity testing, a regulatory requirement for drug discovery and development: Impact of ICH guidelines. Indian J Pharmacol 2002;34:86-99.  Back to cited text no. 8
    
9.
Sardas S. Genotoxicity tests and their use in occupational toxicology as biomarkers. Indoor B E 2005;14:521-5.  Back to cited text no. 9
    
10.
Rivero MT, Vázquez-Gundín F, Goyanes V, Campos A, Blasco M, Gosálvez J, et al. High frequency of constitutive alkali-labile sites in mouse major satellite DNA, detected by DNA breakage detection-fluorescence in situ hybridization. Mutat Res 2001;483:43-50.  Back to cited text no. 10
    
11.
De Méo M, Laget M, Castegnaro M, Duménil G. Genotoxic activity of potassium permanganate in acidic solutions. Mutat Res 1991;260:295-306.  Back to cited text no. 11
    
12.
Olive PL, Banáth JP, Fjell C. DNA strand breakage and DNA structure influence staining with propidium iodide using the alkaline comet assay. Cytometry 1994;16:305-12.  Back to cited text no. 12
    
13.
Uhl M, Helma C, Knasmüller S. Evaluation of the single cell gel electrophoresis assay with human hepatoma (HepG2) cells. Mutat Res 2000;468:213-25.  Back to cited text no. 13
    
14.
Etebari M, Zolfaghari B, Jafarian-Dehkordi A, Rakian R. Evaluation of DNA damage of hydro-alcoholic and aqueous extract of echium amoenum and nardostachys jatamansi. J Res Med Sci 2012;17:782-6.  Back to cited text no. 14
    
15.
Rabbani M, Sajjadi SE, Sadeghi M. Chemical composition of the essential oil from kelussia odoratissima Mozaff. and the evaluation of its sedative and anxiolytic effects in mice. Clinics (Sao Paulo) 2011;66:843-8.  Back to cited text no. 15
    
16.
Etebari M, Ghannadi A, Jafarian-Dehkordi A, Ahmadi F. Genotoxicity evaluation of aqueous extracts of Co-toneaster discolor and Alhagi pseudalhagi by com-et assay. J Res Med Sci 2012;17:S238-42.  Back to cited text no. 16
    
17.
Tavakoli M, Bateni E, Rismanchian M, Fathi M, Doostmohammadi A, Rabiei A, et al. Genotoxicity effects of nano bioactive glass and Novabone bioglass on gingival fibroblasts using single cell gel electrophoresis (comet assay): An in vitro study. Dent Res J (Isfahan) 2012;9:314-20.  Back to cited text no. 17
    
18.
Stoyanova II, Gulubova MV. Mast cells and inflammatory mediators in chronic ulcerative colitis. Acta Histochem 2002;104:185-92.  Back to cited text no. 18
    
19.
Mostafavi SH, Fazilati M, Mostafavi SA, Vahhabi MR, Mostafavi F, Omidvarinia S, et al. Hepatotoxicity of Dorema aucheri (Bilhar) in albino mice. Arch Iran Med 2013;16:530-2.  Back to cited text no. 19