GC-MS and HPLC supported phytochemical analysis of watercress and the protective role against paraben toxicity.

In this study, the phytochemical content of Nasturtium officinale R. Br. (watercress) leaf extract (Noex) and its protective effects against paraben toxicity were investigated. GC-MS and HPLC analyses were performed to determine the phytochemical content. Paraben toxicity and protective properties of Noex were investigated with the Allium test, and 6 different groups were formed for this purpose. Toxicity in each group was investigated by using physiological, cytogenetic, biochemical, and anatomical parameters. DNA-paraben interaction was investigated with spectroscopic analysis for the genotoxicity mechanism. As a result of the study, paraben (500 mM) caused a regression in the physiological parameters related to germination in Allium cepa L. bulbs. Paraben caused a 43.3% reduction in mitotic index (MI) rates compared to control, which is likely the reason for the decrease in germination-related parameters. With the application of paraben in root tip cells, the frequency of micronucleus (MN) and chromosomal aberrations (CAs) increased and a high genotoxic effect was observed. Paraben promoted CAs such as fragment, sticky chromosome, bridge, unequal distribution of chromatin, and irregular mitosis. It also caused anatomical damage in the form of epidermis cell damage, flattened cell nucleus, cortex cell damage, cortex cell walls thickening, and unclear vascular tissue in root tip meristem cells. Paraben-DNA interaction was caused by bathochromic and hypochromic shifts in the UV spectrum of DNA, indicating the intercalation mode of interaction. Paraben also caused an increase in malondialdehyde (MDA) levels, a decrease in glutathione (GSH) levels, and abnormalities in antioxidant enzyme levels (superoxide dismutase = SOD and catalase = CAT), thereby disrupting the antioxidant/oxidant dynamics in the cell. The basis of physiological, cytological, and genetic abnormalities was attributed to the oxidative stress in the cell. Administration of Noex produced a dose-dependent incremental improvement in paraben-induced abnormalities. The increase in GSH levels and the decrease in MDA levels observed as a result of the Noex application contributed to the restoration of antioxidant/oxidant balance, and this improvement was also reflected in other parameters. Application of 200 mg/L Noex provided a 24.2% improvement in the MI rate reduced by paraben, and accordingly, an increase in germination parameters was observed. Similarly, the frequencies of MN and CAs, which are signs of genotoxicity, decreased with the Noex application. As a result of the phytochemical analysis of Noex with HPLC and GC-MS, the presence of strong antioxidant and antimutagenic substances such as rutin, coumaric acid, ferrulic acid, L-serine, L-proline, and phytol were determined in Noex structure. The curative effects of Noex against paraben toxicity can be attributed to these active ingredients.

Genotoxic effects and molecular docking of 1,4-dioxane: combined protective effects of trans-resveratrol.

In this study, the protective effects of trans-resveratrol (t-resv) against 1,4-dioxane-induced toxicity in meristematic cells were investigated. For this purpose, Allium test was used and the alterations in all experimental groups were examined by using physiological, cytogenetic, biochemical, and anatomical parameters. In order to elucidate the toxicity mechanism, interactions of 1,4-dioxane and intracellular antioxidant molecules were investigated by molecular docking. As a result of the analysis, it was determined that 1,4-dioxane causes serious abnormalities in Allium cepa meristematic cells. In 1,4-dioxane-treated group, germination percentage was regressed 1.6 times, root length was reduced 12.7 times, and weight gain was decreased 7.7 times compared to control group. T-resv administration with 1,4-dioxane resulted in an improvement in physiological parameters and reduced the relative injury rate from 0.4 to 0.16. Mitotic index (MI), micronucleus (MN), and chromosomal abnormality (CAs) frequencies were investigated as cytogenetic parameters. 1,4-Dioxane decreased MI index, and increased CAs and MN frequency. In addition, it was determined by the comet test that 1,4-dioxane caused deterioration in DNA integrity. T-resv treatment was found to cause a dose-dependent improvement in genotoxic effects. Changes in the antioxidant system in all experimental groups were determined by measuring malondialdehyde (MDA) and glutathione (GSH) levels, superoxide dismutase (SOD), and catalase (CAT) enzyme activities. 1,4-Dioxane caused alterations in all tested parameters, causing deterioration in the oxidant/antioxidant balance in the cell. A 200-mg/L t-resv+1,4-dioxane treatment caused a 1.9-fold decrease in MDA level which is indicator of lipid peroxidation compared to only 1,4-dioxane-treated group. The mechanism of the disruption in antioxidant/oxidant dynamics and genetic integrity was elucidated by molecular docking analysis of 1,4-dioxane with antioxidant molecules and DNA. In 1,4-dioxane treatment group, anatomical changes such as cell deformation, flattened cell nucleus, and thickening of cortex cell wall were observed. The frequency of these changes decreased with t-resv administration. As a result, it was determined that 1,4-dioxane caused a versatile toxicity in A. cepa meristematic cells, while t-resv was found to have a dose-dependent protective feature against 1,4-dioxane-induced toxicity.

A versatile model for investigating the protective effects of Ceratonia siliqua pod extract against 1,4-dioxane toxicity.

In this study, the toxic effects of 1,4-dioxane, a common contaminant, and the protective property of Ceratonia siliqua L. pod extract (Cspe) against this toxicity are aimed to be demonstrated with a versatile model. For this purpose, Allium toxicity test was used and six different experimental groups were formed. While the control group was germinated in tap water, the application groups were germinated in mediums containing 750 mg/L Cspe, 1500 mg/L Cspe, 100 mg/L 1,4-dioxane, 750 mg/L Cspe+100 mg/L 1,4-dioxane, and 1500 mg/L Cspe+100 mg/L 1,4-dioxane. Each group was germinated in related solution for 72 h and alterations in physiological, biochemical, genetic, and anatomical parameters were investigated. Germination percentage, relative injury rate, root length, and weight gain parameters were examined as physiological parameters, and no significant difference was observed in the control group and only-Cspe-treated groups. In groups treated with 100 mg/L 1,4-dioxane, germination percentage, root length, and weight gain were significantly decreased, and the relative injury rate reached the highest value as 0.48. It was determined that all physiological parameters improved in the groups where Cspe and 1,4-dioxane treated together, and the relative injury rate decreased to 0.22 in the group treated with 1500 mg/L Cspe+1,4-dioxane. Genotoxic effects were tested by the micronucleus and chromosomal abnormality frequency, and statistically insignificant micronucleus formation was found in control group and Cspe-treated groups. Micronucleus frequency were found to be 58.00 ± 12.12 and 31.00 ± 07.38 in 1,4-dioxane and 1500 mg/L Cspe+1,4-dioxane-treated groups, respectively. This result showed that the application of 1500 mg/L Cspe had a 46.5% reduction in the frequency of 1,4-dioxane-induced micronucleus and had a protective effect on genomic integrity. It has been found that 1,4-dioxane application induces lipid peroxidation and increases malondialdehyde level 4.5 times compared with control group. Oxidative stress, which was proved by increased malondialdehyde levels in 1,4-dioxane-treated group caused induction of superoxide dismutase and catalase enzymes, and it was determined that enzyme activities increased by 1.99 and 4.9 times, respectively, compared with the control group. Cspe treatment with 1,4-dioxane caused a significant decrease in malondialdehyde level, superoxide dismutase, and catalase enzyme activities, indicating that oxidative stress formation in the cells was repressed. Abnormalities such as cell deformation, cell wall thickening, and flattened cell nuclei were seen in 1,4-dioxane-treated group in the cross sections of root tips, and the frequency of these abnormalities decreased with Cspe application. As a result, it was determined that 1,4-dioxane caused a versatile toxicity in the test material Allium cepa, whereas Cspe application had a dose-dependent protective feature against toxicity in all tested parameters.