The ubiquity of pollutants, such as agrochemicals and heavy metals, constitute a serious risk to human health. To evaluate the induction of DNA damage and programmed cell death (PCD), root cells of Allium cepa and Vicia faba were treated with two organophosphate insecticides (OI), fenthion and malathion, and with two heavy metal (HM) salts, nickel nitrate and potassium dichromate. An alkaline variant of the comet assay was performed to identify DNA breaks; the results showed comets in a dose-dependent manner, while higher concentrations induced clouds following exposure to OIs and HMs. Similarly, treatments with higher concentrations of OIs and HMs were analyzed by immunocytochemistry, and several structural characteristics of PCD were observed, including chromatin condensation, cytoplasmic vacuolization, nuclear shrinkage, condensation of the protoplast away from the cell wall, and nuclei fragmentation with apoptotic-like corpse formation. Abiotic stress also caused other features associated with PCD, such as an increase of active caspase-3-like protein, changes in the location of cytochrome C (Cyt C) toward the cytoplasm, and decreases in extracellular signal-regulated protein kinase (ERK) expression. Genotoxicity results setting out an oxidative via of DNA damage and evidence the role of the high affinity of HM and OI by DNA molecule as underlying cause of genotoxic effect. The PCD features observed in root cells of A. cepa and V. faba suggest that PCD takes place through a process that involves ERK inactivation, culminating in Cyt C release and caspase-3-like activation. The sensitivity of both plant models to abiotic stress was clearly demonstrated, validating their role as good biosensors of DNA breakage and PCD induced by environmental stressors.

Bisphenol A (BPA) is an endocrine disruptor whose ubiquitous presence in the environment has been related with impairment of male reproduction. BPA can cause both transcriptomic and epigenetic changes during spermatogenesis. To evaluate the potential effects of male exposure to BPA, adult zebrafish males were exposed during spermatogenesis to doses of 100 and 2000 μg/L, which were reported in contaminated water bodies and higher than those allowed for human consumption. Fertilization capacity and survival at hatching were analysed after mating with untreated females. Spermatogenic progress was analysed through a morphometrical study of testes and apoptosis was evaluated by TUNEL assay. Testicular gene expression was evaluated by RT-qPCR and epigenetics by using ELISA and immunocytochemistry. In vitro studies were performed to investigate the role of Gper. Chromatin fragmentation and the presence of transcripts were also evaluated in ejaculated sperm. Results on testes from males treated with the highest dose showed a significant decrease in spermatocytes, an increase in apoptosis, a downregulation of ccnb1 and sycp3, all of which point to an alteration of spermatogenesis and to meiotic arrest and an upregulation of gper1 and esrrga receptors. Additionally, BPA at 2000 μg/L caused missregulation of epigenetic remodelling enzymes transcripts in testes and promoted DNA hypermethylation and H3K27me3 demethylation. BPA also triggered an increase in histone acetyltransferase activity, which led to hyperacetylation of histones (H3K9ac, H3K14ac, H4K12ac). In vitro reversion of histone acetylation changes using a specific GPER antagonist, G-36, suggested this receptor as mediator of histone hyperacetylation. Males treated with the lower dose only showed an increase in some histone acetylation marks (H3K14ac, H4K12ac) but their progeny displayed very limited survival at hatching, revealing the deleterious effects of unbalanced paternal epigenetic information. Furthermore, the highest dose of BPA led to chromatin fragmentation, promoting direct reproductive effects, which are incompatible with embryo development.

This study examines whether selected functional food and medicinal plants can mitigate the adverse effects of xylene on ovarian cells. The influences of xylene (0, 10, 100, or 1000 ng/mL), buckwheat (Fagopyrum esculentum), rooibos (Aspalathus linearis), vitex (Vitex agnus-castus), extracts (10 μg/mL each), and a combination of xylene with these plant additives on cultured porcine ovarian granulosa cells are compared. Cell viability, proliferation (PCNA accumulation), apoptosis (accumulation of bax), and release of progesterone (P4) and estradiol (E2) were analyzed by the trypan blue tests, quantitative immunocytochemistry, and enzyme-linked immunosorbent assays, respectively. Xylene suppressed all measures of ovarian cell function. Rooibos prevented all of xylene’s effects, whereas buckwheat and vitex prevented four of five of the analyzed effects (buckwheat prevented xylene influence on viability, PCNA, bax, and E2; vitex prevented xylene action on viability, PCNA, and P4 and E2). These observations show that xylene has the potential to suppress ovarian cell functions, and that buckwheat, rooibos, and vitex can mitigate those effects, making them natural protectors against the adverse effects of xylene on ovarian cells.

We evaluated the influence of the oil-related environmental contaminant benzene (0, 10, 100, or 1000 ng/mL) alone and in combination with apigenin, daidzein, or rutin (10 μg/mL each) on viability; proliferation (accumulation of proliferating cell nuclear antigen); apoptosis (accumulation of Bax); and release of progesterone (P), testosterone (T), and estradiol (E) in cultured porcine ovarian granulosa cells. Cell viability; proliferation; apoptosis; and release of P, T, and E have been analyzed by the trypan blue test, quantitative immunocytochemistry, and ELISA, respectively. Benzene did not affect apoptosis, but reduced ovarian cell viability and P and E release, and promoted proliferation and T output. Apigenin did not affect cell viability, but stimulated proliferation and T and E release, and inhibited apoptosis and P secretion. It prevented and reversed the action of benzene on proliferation and P and T release, and induced the inhibitory action of benzene on apoptosis. Daidzein promoted cell viability, proliferation, P release, but not apoptosis and T or E release. Daidzein induced the stimulatory effect of benzene on T, without modifying other effects. Rutin administered alone reduced cell viability and apoptosis, and promoted cell proliferation. Furthermore, rutin prevented and reversed the effect of benzene on proliferation and P and E release. These observations suggest the direct action of benzene and plant polyphenols on basic ovarian cell functions, and the ability of apigenin and rutin, but not of daidzein, to prevent benzene effects on the ovary.

We aimed to examine the influence of benzene and of three dimethyl sulfoxide (DMSO) plant extracts—buckwheat (Fagopyrum Esculentum), rooibos (Aspalathus linearis), and vitex, (Vitex Agnus-Castus), and the combination of benzene with these three plant extracts on basic ovarian cell functions. Specifically, the study investigated the influence of benzene (0, 10, 100, or 1000 ng/mL) with and without these three plant additives on porcine ovarian granulosa cells cultured during 2 days with and without these additives. Cell viability, proliferation (accumulation of proliferating cell nuclear antigen, PCNA), apoptosis (accumulation of Bcl-2-associated X protein , bax), and the release of progesterone (P) and estradiol (E) were analyzed by the Trypan blue test, quantitative immunocytochemistry, and enzyme-linked immunosorbent assay, respectively. Benzene reduced cell viability, as well as P and E release. Plant extracts, given alone, were able directly promote or suppress ovarian cell functions. Furthermore, buckwheat and rooibos, but not vitex prevented the inhibitory action of benzene on cell viability. Buckwheat induced the stimulatory action of benzene on proliferation. Rooibos and vitex promoted benzene effect on cell apoptosis. All these plant additives were able to promote suppressive action of benzene on ovarian steroidogenesis.These observations show that benzene may directly suppress ovarian cell viability, P, and E release and that buckwheat, rooibos, and vitex can directly influence ovarian cell functions and modify the effects of benzene—prevent toxic influence of benzene on cell viability and induce stimulatory action of benzene on ovarian cell proliferation, apoptosis, and steroidogenesis. The observed direct effects of benzene and these plants on ovarian cells functions, as well as the functional interrelationships of benzene and these plants, should be taken into account in their future applications.