BIYOLOGY

This study investigated the in vitro effects of sodium fluoride (NaF) on cytotoxicity and oxidative stress in TM3 Leydig cells. Cytotoxicity was determined with cell viability and proliferation rate, and lactate dehydrogenase leakage assay. Oxidative damage was measured by increasing lipid peroxidation product, reactive oxygen species (ROS) and decreasing cellular enzymatic and non-enzymatic antioxidants. Leydig cells were exposed to two different concentrations of NaF (2ppm and 16ppm) for 24h, 48h and 72h which are considered to be the lowest and highest fluoride concentrations based on daily intake of drinking water in Turkey. The present study showed that cell viability and proliferation decreased and oxidative stress increased in NaF-exposed Leydig cells in a concentration-and time-dependent manner. The findings suggest that fluoridetreatment can cause damage in Leydig cells, especially in antioxidant system of the cells. The results also proved that oxidative stress plays an important role in impairments on Leydig cell function.

Polychlorinated biphenyls (PCBs) are ubiquitous and persistent environmental contaminants that disrupt endocrine function in biological systems, especially in the male reproductive system. Previous studies on the reproductive toxicity of PCBs have focused on the impairment of spermatogenesis, disruption of steroidogenesis, decreased sperm number, and infertility. Aroclor 1242 is a commercial mixture with an average of 42% chlorine by weight. The purpose of the present study was to elucidate the hazardous effects of Aroclor 1242 on Leydig cells through an evaluation of cell viability, lipid peroxidation, hydroxyl radicals, H2O2production, antioxidant enzymes, and steroidogenic enzymes. Leydig cells were exposed to Aroclor 1242 for 24 h under basal and luteinizing hormone-stimulated conditions at different concentrations (ranging from 10−16M to 10−6M). After incubation, Leydig cells were measured for cell viability, lipid peroxidation, reactive oxygen species (hydroxyl radical and H2O...

The present study was aimed at determining the oxidative damage caused by sodium arsenite in 3T3 fibroblast cells and the possible protective role of curcumin (Cur) against sodium arsenite toxicity. Embryonic fibroblast cells were exposed to sodium arsenite (0.01, 0.1, 1, and 10 μM) in the presence and absence of Cur (2.5 μM) for 24 hours. Cell viability, cytotoxicity, lipid peroxidation, hydroxyl radical, hydrogen peroxide, antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase, and glutathione-S-transferase) and expression levels of antioxidant genes (superoxide dismutase, catalase, and glutathione peroxidase) were measured in embryonic fibroblast cells. Results demonstrated that sodium arsenite directly affects antioxidant enzymes and genes in 3T3 embryonic fibroblast cells and induces oxidative damage by increasing the amount of hydrogen peroxide, hydroxyl radical, and lipid peroxidation in the cell. Furthermore, the study indicated that Cur might be a potential ameliorative antioxidant to protect the fibroblast cell toxicity induced by sodium arsenite.

Penicillium citrinum-derived mycotoxin citrinin (CTN) is known to be a toxic agent for humans and animals. Previous studies have shown that CTN leads to toxicity in many biological systems; however, a limited number of studies have been performed to demonstrate the harmful effects of CTN on the male reproductive system. In the present study, the effects of CTN on cytotoxicity and apoptosis were examined in Sertoli cells as a model. Sertoli cells were treated with eight different CTN concentrations (from 0 up to 200 mM, for 6-72 h). Toxic potential of CTN was estimated by measuring metabolic activity (MTT test), DNA synthesis (BrdU test), and cell membrane damage (LDH test) as well as apoptosis and necrosis (via staining with propidium iodide and Hoechst 33342). The results showed that CTN significantly decreased the cell viability and cell proliferation, increased cytotoxicity, apoptosis, and necrosis in a concentration-dependent manner. Furthermore, CTN showed cytotoxicity in Sertoli cells with an IC50 value of 116.5 mM for 24 h. In conclusion, these findings clearly showed that, CTN affects Sertoli cells even at low concentrations. Thus, as a result of the damage of CTN shown in Sertoli cells, it can be deduced that CTN may also have detrimental effects on the testes.

Acrolein is a highly reactive unsaturated organic molecule and has harmful effects on human health. Acrolein is generally formed in heat-treated foods above 150°C, as well as in the combustion of gasoline, wood industry, plastic waste, and tobacco smoke. In this study, the effects of acrolein on genotoxicity in Leydig cells and the underlying mechanisms are aimed to be clarified. In addition, the toxicogenomic profile of acrolein was studied in terms of both apoptosis and steroidogenesis. Real-time PCR and ELISA tests were used to analyses of steroidogenic endpoints. Apoptosis was evaluated with double fluorescence staining and gene expression analyses of related genes. Comet assay was used to determine the genotoxicity. The results showed that acrolein caused concentration-dependent inhibition on cell viability at 8 μM and above concentrations, decreased testosterone production at 13.6 and 19.7 μM concentrations, and suppressed expression levels of genes that play an important role in steroidogenic pathway. Furthermore, acrolein downregulated expression of anti-apoptotic Bcl2 gene and upregulated expression of proapoptotic Bax, Casp3, and Trp53 gene after 24-h treatment in 7.4, 13.6, and 19.7 μM acrolein-exposed Leydig cells. The results of comet assay showed that acrolein significantly induced tail length, tail % DNA, and Olive tail moment. Overall, it was concluded that acrolein-induced cell damage in Leydig cells may be due to formation of genetic damage in steroidogenesis and apoptosis.