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. 2019 Mar 16;5(3):e01337. doi: 10.1016/j.heliyon.2019.e01337

Herbal medicine against genotoxicity of dimethoate, an insecticide, in mammalian somatic cells

Halit Kızılet a, Bilal Yilmaz b, Handan Uysal c,∗
PMCID: PMC6423489  PMID: 30923767

Abstract

In this study, the genotoxic effects of dimethoate (DIM) were investigated with the in vitro micronucleus test in human peripheral lymphocytes. The ethanol extracts of Rosa canina and Salvia lavandulifolia were used to remove possible genotoxic effects of these substances. For this purpose, different concentrations (0.5-1-2 μg/mL) of dimethoate, DIM + RCeta and DIM + SLeta (1:1 v/v) application groups were prepared and applied to the blood culture. The obtained data were compared with the negative control group that was prepared with dimethyl sulfoxide (DMSO) as solvent and a well-known genotoxic effects of ethyl methanesulfonate (EMS) as positive control group. It was observed in lymphocyte cells that the frequency of MN considerably increased depending on the increasing dose of DIM whereas the nuclear division index (NBI)decreased according to the control group, especially in the last concentration (2 μg/mL). But, as the MN frequency decreased, NBI values approached to control group with 2μg/mL DIM + RCeta and 2μg/mL DIM + SLeta according to DIM application group (P < 0.05). Additionally, RCeta and SLeta were analyzed by gas chromotography-mass spectrometry (GC-MS).

Keywords: Toxicology, Genetics

1. Introduction

The farmed areas of agriculture in the world is fixed at 1.400.000 ha from 1950 to the present day, while the human population has from 2.600.000 reach to 7.600.000 since 1950 to 2017 (Info, 2017; UN, 2017). The increasing population together with the environmental pollution in the world is also increasing in different ecosystems. Especially in agricultural areas, plants are under the threat of microorganisms, harmful insects, rodents and weeds from the sowing to harvest time and from storage to processing stage. Pesticides are biologically active chemical substances used against harmful organisms to increase the amount of product that can be taken from the unit area. For this reason, pesticides and insecticides –a sub-group of pesticides-have been developed against plant diseases and pests to increase the yield obtained from the unit area in agriculture. These substances have been shown to cause destruction of useful organisms and endangering genetic diversity as they are used more and unconsciously against pests, weeds and diseases every day for the purpose of agricultural struggle (Ozkaraova Gungor, 2008).

Organophosphorus compounds (sub-group of insecticides) may induce oxidative stress leading to the generation of free radicals (Sharmaa et al., 2005). Dimethoate (O,O-dimethyl-S-N-methylcarbamoylmethyl)-phosphorodithioate) [CAS registry no.60-51-5] is an organophosphorus insecticide with a contact and systemic action. It is widely used against a broad spectrum of insects, mites and houseflies. The main risk groups of higher-dose dimethoate exposure are the producers, pesticide workers, farm owners and households.

In many studies, it has been determined that insecticides cause genomic instability, which leads to pathological cases and even aging with various diseases including cancer (Soltani et al., 2009). In order to prevent this situation, studies related with the determination of antimicrobial, antioxidant, antimutagenic, anticancerogenic and even antiaging characteristics of natural compounds originated from plants containing different phytochemicals are being continued (Arora et al., 2002; Simopoulos, 2004; Uysal et al., 2009a, 2015, 2009b; Kızılet et al., 2013; Kasımoğlu and Uysal, 2014). According to Jain et al. (2007), although the science of medicine has grown tremendously in the 20th century, the use of plants still continues in traditional medicine/alternative medicine.

A knowledge of the chemical constituents of rosehip (Rosa canina) and sage (Salvia lavandulifolia) (which grows naturally in Turkey) are desirable not only for the discovery of therapeutic agents, but also because such information may be of great value in disclosing new sources of economic phytocompounds for the synthesis of complex chemical substances and for discovering the actual significance of folkloric remedies. Hence a thorough validation of the herbal drugs has emerged as an emerging issue of interest emphasizing and prioritizing the standardization of the natural drugs and products because several of the phytochemical have complementary and overlapping mechanism of action. Mass spectrometry coupled with chromatographic separations such as gas chromatography (GC-MS) is normally used for direct analysis of components existing in traditional medicines and medicinal plants.

In this study, the genotoxicity of dimethoate insecticide was determined by micronucleus technique in human peripheral lymphocyte cells. In addition, these genotoxic effects were eliminated by ethanol extracts of R. canina (RCeta) and S. lavandulifolia (SLeta) Also, in this study, thechemical contents of the R. canina and S. lavandulifolia plants were determined by GS-MS method.

2. Materials and methods

2.1. Preparation of plant extracts

R. canina and S. lavandulifolia used in this study were collected from Palandöken Mountain of Erzurum province, and Aşkale district, Koşapınar village, respectively. All above-ground structures (stem, leaf, flower etc.) of S. lavandulifolia and just fruits of R. canina had been dried and ground in a cool and clean environment without exposure to direct light. 50 g of ground plant was extracted with 150 mL of ethanol. The ethanol and plant mixture were passed through filter paper after standing at room temperature for 24 hours. In order to increase the amount of solution, this procedure was repeated three times. The mixed or unified filtrates were concentrated with a Soxhlet extractor at 50 °C. 2 μg/mL ethanol extracts (SLeta and RCeta) were dissolved in 2% dimethyl sulfoxide (DMSO) during the application.

2.2. GC-MS system

Chromatographic analysis were carried out on an Agilent 7820A gas chromatography system equipped with 5977 series mass selective detector, 7673 series autosampler and chemstation (Agilent Technologies, Palo Alto, CA). HP-5 MS column with 0.25 μm film thickness (30 m × 0.25 mm I.D., USA) was used for separation. The temperatures of the inlet, transfer line and detector were 250, 250 and 300 °C, respectively.

2.3. GC-MS conditions

Different temperature programs were investigated for GC-MS method. The end of this investigation, the temperature program of the GC/MS was as follows: initial temperature was 50 °C, held for 1 min, increased to 100 °C at a rate of 20 ºC/min held for 1 min, increased to 180 °C at a rate of 10 ºC/min held for 1 min, increased to 220 °C at rate of 5 ºC/min held for 5 min, and finally to 300 °C at a rate of 10 min and held for 5.5 min. The injector volume was 1 μl in splitless mode and the carrier gas was helium at a flow rate of 1 ml/min.

2.4. Identification of components

The spectrum of the unknown component was compared with the spectrum of the component stored in the National Institute of Standards and Technology Library Version (2005), Software, Turbomass 5.2. The components were identified by comparing linear Kovats retention index and mass spectra with those obtained from the MS library. Interpretation on mass spectrum GC-MS was conducted using the database on National Institute Standard and Technology having more than 62,000 patterns. The relative percentage amount of each component was calculated by comparing its average peak area to the total areas. The name, molecular weight and structure of the components of the test materials were ascertained.

2.5. Donors with peripheral blood assays

The study involved control and experimental group composed of 3 healthy volunteers with the age of 25–30 years. None of the individuals in the study were habitual smokers or drinkers, and had not been exposed to any known toxic agents, drugs or X rays.

The ethics committee permission for the study was received from Erzurum Regional Training and Research Hospital Local Ethics Committee (Number: 37732058-53/2467/ BEAH KAEK 2015/9-67) and the rules of the committee were followed during the investigations. Written informed consent was obtained from all patients who participated in this study.

2.6. In vitro micronucleus (MN) test

A newly modified version of micronucleus test developed by Fenech (2000) and Kirsch-Volders et al. (1997) was used in this study. Preliminary studies were conducted to determine the DIM applicationdoses in which cell division was not blocked and 0.5, 1.0 and 2.0 μg/mL were determined as doses used throughout the study. Enough number of divided cells could not reach at higher dosesthan these application groups. In our study, distilled water and 2% DMSO, the solvent of DIM, were used as negative controls and 10 mM ethyl metansulfonate (EMS) was used as a positive control apart from DIM application groups. In the second part of the study, another experimental setup with 2.0 μg/mL DIM + RCeta and 2.0 μg/mL DIM + SLeta (1:1 v/v equal amount of insecticide and plant extract) application groups were prepared. Experiments were repeated three times with different donors for each treatment group. All treatment groups were incubated for 72 hours with blood from appropriate donors. Two nucleated cells (binucleat) were obtained with cytochalasin-B added at 48th hour in each tubes. At the end of the incubation, prepared smear slides were stained with Giemsaand investigated under 10X40 magnification.

1000 cells were randomly counted and the Nuclear Division Index (NDI) was calculated by determining the ratio of one, two, three and four nuclei cells for each repeat. In addition, 1000 two nucleated (binucleat) cells were counted and MNs were recorded. According to the results, DMSO and EMS compared with distilled water, all DIM applications compared with DMSO, and plant extract + DIM applications compared with 2 μg/mL DIM. SPSS 13.0 program was used for statistical analysis related to MN and NBI values obtained as a result of the studies. Univariate analysis of variance (ANOVA) and Turkey test were applied to compare the data obtained from control and treatment groups.

3. Results

As a result of our experiments, the total MN percentages were found as 0.70 ± 0.38 in the distilled water and 0.83 ± 0.65 in the DMSO. In the EMS positive control group, this value was obtained as 5.63 ± 1.60. The difference between the distilled water and the DMSO control group was insignificant (P > 0.05), while the difference between the EMS positive control and the negative control group distilled water was found to be significant (P < 0.05). NDI values of control groups were obtained as 1.52 ± 0.15 for distilled water group; 1.54 ± 0.17 for DMSO group and 1.29 ± 0.25 for EMS group. When NDI results of the DMSO and EMS groups were compared to those of the distilled water group, it was determined that EMS has a cytotoxic effect (P < 0.05) but DMSO not (P > 0.05).

The results obtained from DIM applications are shown in Table 1. The MN percentages due to DIM applications at 0.5 and 1.0 μg/mL were found as 0.90 ± 0.72 and 0.97 ± 0.63, respectively. Moreover, NDI values were obtained as 1.54 ± 0.20 and 1.54 ± 0.23 respectively. The differences between the MN and NBI values of these two DIM groups and the DMSO control group was non-significant (P > 0.05). The MN percentage and NDI value for 2.0 μg/mL DIM application were obtained as 2.10 ± 0.13 and 1.29 ± 0.32, respectively (Table 1). According to these results, the differences in the MN percentage and NDI values for 2.0 μg/mL DIM application and the DMSO control group were found to be significant (P < 0.05).

Table 1.

Statistical significance of MN and NDI values after the exposure to DIM, DIM + RCeta and DIM + SLeta.

Application groups Concentration (μg/mL) BNC MN within binucleat
 %MN±S.E. NDI±S.E.
(1) (2) (3)
Control Distilled water - 3000 21 - - 0.70 ± 0.38 1.52 ± 0.15
DMSO %2 3000 25 - - 0.83 ± 0.65 1.54 ± 0.17
EMS 10mM 3000 107 16 10 5.63 ± 1.60* 1.29 ± 0.25*
Insecticides application groups DIM 0.5 3000 27 - - 0.90 ± 0.72 1.54 ± 0.20
DIM 1.0 3000 29 - - 0.97 ± 0.63 1.54 ± 0.23
DIM 2.0 3000 63 - - 2.10 ± 0.13* 1.29 ± 0.32*
Insecticides + plant extracts DIM + RCeta 2.0 + 2.0 3000 13 - - 0.43 ± 0.16** 1.35 ± 0.41**
DIM + SLeta 2.0 + 2.0 3000 25 - - 0.83 ± 0.20** 1.38 ± 0.28**
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BNC:Investigated of binucleated cells, MN: micronucleus, NDI: nuclear division index, *:The mean difference is significant at the 0.05 level, **:The mean antioxidant difference is significant at the 0.05 level.

To be able to determine the antigenotoxic effects of the plant ethanol extracts, the application of DIM + RCeta and DIM + SLeta (1:1 v/v) was performed. However, RCeta and SLeta treatments were not performed in the 0.5 and 1.0 μg/mL DIM treated groups, since the MN frequencies were found to be extremely low in these groups (Table 1). On the other hand, the MN frequency values with DIM + RCeta and DIM + SLeta applications were significantly decreased to 0.43 ± 0.16 and 0.83 ± 0.20 with respect to the 2.0 μg/mL DIM exposed group (2.10 ± 0.13) (P < 0.05). Moreover, The NDI values for DIM + RCeta and DIM + SLeta groups were measured as 1.35 ± 0.41 and 1.38 ± 0.28 respectively. The differences in the NDI values of DIM, DIM + RCeta and DIM + SLeta groups were found to be significant (P < 0.05).

The components present in the R. canina and S. lavandulifolia are listed in Tables 2 and 3. 33 components (92.97%) and 17 components (90.63%) were identified RCeta andSLeta, respectively.

Table 2.

Composite composition values for the R. canina ethanol extract.

Peak number Retantion time(min) Component Molecular formula Molecular weight(g/mol) % ratio in total component
1 3.52 Hexanal C6H12O 100.09 0.69
2 4.31 Pentanoic acid C5H10O2 102.07 0.32
3 5.14 2,4-dihydroxy-2,5-dimethyl-3(2h)-furanone C6H8O4 144.04 0.18
4 5.21 Furan, 2-penthyl C9H14O 138.10 0.37
5 5.71 Hexanoic acid C6H12O2 116.08 5.21
6 6.69 Heptanoic acid C7H14O2 130.10 0.77
7 8.13 Octanoic acid C8H16O2 144.11 1.41
8 8.72 2-octanoic acid, trans- C8H14O2 142.10 0.40
9 9.09 Butanedioic acid, hydroxy diethylester C8H14O5 190.08 0.55
10 9.51 Nonanoic acid C9H18O2 158.13 1.44
11 9.64 5-Acetylvaleric acid, methylester C8H14O3 158.09 0.68
12 10.25 2-Methyl-oct-2-enedial C9H14O2 154.10 1.27
13 12.26 7-Methyl-Z-tetradecen-1-ol acetate C17H32O2 268.24 0.94
14 12.48 9-Oxononanoic acid C9H16O3 172.11 1.18
15 13.04 4-amino-1,5-pentandioic acid C7H13NO4 175.08 2.06
16 13.34 Dodecanoic acid C12H24O2 200.18 1.36
17 14.85 Azelaic acid, ethylester C11H20O4 216.14 1.83
18 15.61 Azelaic acid C9H16O4 188.10 5.62
19 16.43 Oleic acid C18H34O2 282.26 0.83
20 18.98 3-deoxyestradiol C18H24O 256.18 0.34
21 19.99 Hexadecanoic acid, ethylester C18H36O2 284.27 15.44
22 21.31 10-octadecenoic acid, methylester C19H36O2 296.27 0.98
23 22.59 Trans-13-octadecenoic acid C18H34O2 282.26 3.01
24 23.21 Oleic acid, ethyl ester C20H38O2 310.29 26.43
25 23.75 Stearic acid C18H36O2 284.27 8.27
26 28.03 Cis-13-Eicosenoic acid C20H38O2 310.29 0.95
27 28.59 Eicosanoic acid C20H40O2 312.30 1.39
28 32.87 7-Methyl-Z-tetradecen-1-olacetate C17H32O2 268.24 0.77
29 32.87 7-Methyl-Z-tetradecen-1-olacetate C17H32O2 268.24 0.91
30 33.89 13-Decosenamide C22H43NO 337.33 1.28
31 34.75 Oleic acid, 3-(octadecyloxy) propyl ester C39H76O3 592.58 1.95
32 35.68 Ethyl iso-allocholate C26H44O5 436.32 0.24
33 39.59 17-pentatriacontene C35H70 490.55 3.91
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Table 3.

Composite composition values for the S. lavandulifolia ethanol extract.

Peak number Retantion time(min) Component Molecular formula Molecular weight(g/mol) % ratio in total component
1 3.37 2-octanol C8H18O 130.14 0.22
2 3.63 Methoxyformamide C2H5NO2 75.03 0.37
3 5.16 2,4-dihydroxy-2,5-dimethyl-3(2H)-furan-3-one C6H8O4 144.04 0.65
4 6.01 Erythritol C4H10O4 122.06 3.28
5 7.65 4H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl C6H8O4 144.04 2.39
6 9.00 6-Acetyl-β-d-mannose C8H14O7 222.07 3.46
7 10.76 10-heptadecen-8-ynoic acid, methyl ester C18H30O2 278.22 0.82
8 11.70 Benzofuran-2-carboxaldehyde C9H6O2 146.04 7.47
9 13.28 Β-hydroxydodecanoic acid C12H24O3 246.17 1.23
10 19.77 Hexadecanoic acid C16H32O2 256.24 8.54
11 19.92 Hexadecanoic acid, ethyl ester C18H36O2 284.27 3.89
12 22.15 1-heptatriacotanol C37H76O 536.59 1.75
13 23.14 Linolenic acid C18H30O2 278.22 26.38
14 30.92 Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl) ethyl ester C19H38O4 330.23 3.24
15 33.33 Octadecanoic acid, 2-hydroxy-1-(hydroxymethyl) ethyl ester C21H42O4 358.31 6.13
16 33.87 13-Docosenamide, (Z)- C22H43NO 337.33 19.84
17 34.11 Ethyl iso-allocholate C26H44O5 436.32 0.98
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4. Discussion and conclusion

The imbalance between the antioxidant defense of the body and the free radical formation, which causes peroxidation of the lipid layer of cells, is called oxidative stress. Oxidative stress causes changes in the membrane fluidity and DNA damages. It has also carcinogenic effects (Singh and Pandey, 1989; Bagchi et al., 1995). As in the current study, the balance between oxidant and antioxidant status in the cell is distrupted due to the insecticides, free radicals increase, and this leadts to oxidative stresses and DNA damages. DNA damage and oxidative stress play an important role in cancer, aging, telomere shortening, various diseases and pathological conditions (Martin-Ruiz et al., 2004; Soltani et al., 2009). Recent studies have reported that the oxidative stress and lipid peroxidation increased due to enhanced free radicals formation in the pesticide poisonings (Oruç and Üner, 2000; Hazarika et al., 2003; Shadnia et al., 2005; Lukaszewicz-Hussain, 2010; Roberts and Reigart, 2013). In the current study, a statistically significant differences in MN percentage and NBI values were obtained between 2.0 μg/mL DIM treated and control groups (P < 0.05). According to these results, it is highly possible that DIM treatment caused oxidative stresses in living systems. For example, it has been reported that DIM led to a chromosomal aberrations in mouse bone marrow, an increased SCE in hamster cells, and it also had mutagenic effects in Drosophila (Athwal and Sandhu, 1985). Gomez-Arroyo et al. (1987) reported that methyl parathion and dimethoate insecticides increased sister chromatid exchanges in human peripheral lymphocytes as compared to the control group, and statistically positive results were observed. Moreover, Maiti and Kar (1997), and Maiti et al., (1996), demonstrated that dimethoate intoxication produced oxidative stress through the generation of free radicals and induced hepatic lipid peroxidation in chicken and mice. According to Ündeğer and Başaran (2005), dimethoate and methyl parathion at 100 and 200 μg/mL significantly increased DNA damage in human lymphocytes. In addition, in vitro exposure to dimethoate induced a concentration-dependent increased sister chromatid exchange (SCE) frequency in toadfish lymphocytes (Ellingham et al., 1986). In another study, after exposure to dimethoate, the level of superoxide dismutase declined while lipid peroxidation, glutathione, induction of micronucleus and DNA damage were increased in C. punctatus as the concentration and exposure time increased (Daoud et al., 2014). It is also found to increase the incidence of numerical but not structural chromosomal aberration (CA) in male Wistar rats (Ündeğer et al., 2000). Moreover, it is reported that dimethoate was mutagenic in Escherichia coli, Saccharomyces cerevisia and Salmonella typhimurium TA 100, but non-mutagenic in Salmonella typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 102 (Mohn, 1973; Bianchi et al., 1994). According to Dogan et al. (2011), it can be concluded that oxidative stress condition evoked by dimethoate could not be responded effectively and genotoxic nature of pesticide was proven by determined clastogenic effect possibly via being an alkylation agent or stimulating the production of reactive species.

In addition, it has been shown that profenofos, an organophosphate insecticide, increased the micronucleus frequency in human peripheral lymphocyte cells and decreased the nuclear division index (0.25, 0.5, 0.75 and 1.0 μg/mL) at different concentrations (Kasımoğlu and Uysal, 2014). In another study of antioxidant enzyme levels with chlorpyrifos-ethyl from organophosphate insecticides, it has been found that the paraoxonase activity and total antioxidant levels decreased while the levels of oxidative stress index and total oxidant increased (Deveci et al., 2015). Gültekin et al. (2000) showed that chlorpyrifos, an organophosphate insecticide, increased the level of malondialdehyde (MDA) lipid peroxidation product in erythrocytes. Similarly, Gültekin et al. (2001) reported that chlorpyrifos-ethyl increased lipid peroxidation in rats and the antioxidant defense decreased together with increased oxidative stress. Many studies on plant-derived natural compounds demonstrated that plant compounds exhibit a protective activity against genotoxicity caused by oxidative stress (Plazar et al., 2008). Medicinal plants are a potential source of antioxidants and ROS scavenging molecules (Arora et al., 2005) and contain high amount of diverse vitamins and minerals. Many studies over the past three decades have focused on the assessment of anticarcinogenic and antimutagenic activities of plants. One of these is the rosehip fruit, which contains plenty of various vitamins including vitamin C, B1, B2, E, K and P (permeability vitamins) and carotene. It is also rich in mineral substances and flavonoids (Gao et al., 2000; Hvattum, 2002; Demir et al., 2014). The water extracts of Salvia species, the other plant used in the current study, are known to be rich in rosmarinic acid and luteolin-7-glucoside (Ramos et al., 2010). In our study, the ethanol extracts of R. canina and S. lavandulifolia were applied as much as 2 μg/mL DIM and the results were found to be important for their antigenotoxicity (P < 0,05). These obtained findings indicated that the oxidative damages caused by DIM treatment are eliminated by the free radical scavenging activity of SLeta and RCeta. Similar studies in the literature reported that there was a strong association between the phenolic compounds and the antioxidant activity in R. canina, S. lavandulifolia and many plant species.

In a study done by Kılıçgün (2008), it was determined that R. canina inhibited liver damage in rats even at low concentrations. The ascorbic acid, which was found as large amount of R. canina, was demonstrated to reduce the genotoxic effects of EMS, MMS (methyl methanesulfonate) and ENU (N-nitroso N-ethylurea) in D. melanogaster (Kaya, 2003). Moreover, the other compund, beta carotene in R. canina was shown to decrease the genotoxic effects of doxorubicin in this fruit fly (Dias et al., 2009). It was found that the ethanol and water extracts of R. canina reduced the genetic damages in the human lymphocyte cells induced by cypermethrin and fenvalerate insecticides exposures and in the D. melanogaster induced by EMS exposure (Kızılet et al., 2013; Kasımoğlu and Uysal, 2014). Ramos et al. (2010) demonstrated that the extracts of S. officinalis, S. fruticosa, and S. lavandulifolia and their major components; rosmarinic acid and luteolin-7-glucoside protected the DNA of Caco-2 and HeLa cells exposed to oxidative agents against oxidative DNA damage and stimulated DNA repair. Eidi and Eidi (2009) investigated the antidiabetic effects of S. officinalis leaves on normal and diabetic mice. It was shown that the ethanol extract of these leaves led to a reduction in the amount of serum glucose, triglyceride and total cholesterol in diabetic mice, while it had no effects in normal mice. In another study performed by Uysal et al. (2015), it was observed that the methanol extract of Echium amoenum reduced the genotoxic effects of EMS.

In RCeta sample (Table 2), 92.97% of the total extract was identified, predominating oleic acid (29.21%), hexadecanoic acid (15.44%), stearic acid (8.27%), azelaic acid (7.45%) hexanoic acid (5.21%), octadecenoic acid (3.99%) and pentatriacontene (3.90%). In SLeta sample (Table 3), 90.63% of the total extract content was identified and the main components were linolenic acid (26.38%), docosenamide (19.84%), hexadecanoic acid (15.67%), benzofuran-2-carboxaldehyde (7.47%), octadecanoic acid (6.13%), 6-acetyl-β-d-mannose (3.46%) and erythritol (3.28%). Major fatty acids has biological activities such as being anti-inflammatory, anti-androgenic, cancer preventing, dermatigenic, hypocholesterolemic, anaemiagenic, insectifuge as well as being a 5-alpha reductase inhibitör (Duke, 1998). n-hexadecanoic acid has been reported to show selective toxicity to human leukemic cells as well as anti tumor activity and is suggested to be a lead compound in anti cancer drugs (Harada et al., 2002).

These results MN demonstrate that DIM can cause genotoxic effects even at very low doses. Since genotoxicity induced in somatic cells is highly likely to occur gametically, plants with medical prescription as alternative genomic preservatives should be taken into consideration in the daily nutritional regime. As a result, we can say that theseplant components, belonging to R. caninaand S. lavandulifolia, protect the cells against oxidative DNA damage and stimulate DNA repair. Our obtained results were consistent with the similar studies on plant extracts. According to the results obtained from our study, it can be inferred that the scavenging effects of R.canina ve S. lavandulifolia on the DIM was arisen from the antioxidative properties of their compounds.

Declarations

Author contribution statement

Halit Kızılet, Bilal Yilmaz, Handan Uysal: Conceived and designed the experiments; Performed the experiments; Analyzed and interpreted the data; Contributed reagents, materials, analysis tools or data; Wrote the paper.

Funding statement

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Competing interest statement

The authors declare no conflict of interest.

Additional information

No additional information is available for this paper.

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