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

Abstract

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 (500mM) 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 200mg/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.