Toxic effect of antidepressants on male reproductive system cells: evaluation of possible fertility reduction mechanism.

Depression is acknowledged as a major public health problem. Pharmacological treatment may cause adverse drug reactions and sexual side effects. At the same time, the knowledge of the molecular mechanisms associated with antidepressant-mediated toxicity to reproductive cells is fragmentary. The aim of this study was the multilevel evaluation of the potential toxicity of several antidepressants or antipsychotic drugs (amitriptyline, 10 µM; escitalopram, 30 µM; fluoxetine, 5 µM; imipramine, 20 µM; mirtazapine, 150 µM; olanzapine, 40 µM; reboxetine, 30 µM; venlafaxine, 250 µM) on the cells of the spermatogenesis pathway. Effects of various drugs were monitored by several methods including mitochondrial activity MTT test, fluorescent staining, real-time PCR, morphology analysis, immunofluorescence, and Western blots. Obtained results suggest the concentration- and the time-dependent cytotoxic effect. The molecular mechanism of cytotoxic effect is mediated by disturbances in the redox balance (increased production of reactive oxygen species and reactive nitrogen species), failure of enzymatic and non-enzymatic cell protection mechanisms (glutathione system, nuclear factor-κB and fibroblast growth factor 2-mediated pathways), and impairment of mitochondrial functions. In addition, we provide for the first time, to our knowledge, evidence that antidepressant treatment may contribute to spindle apparatus assembly defects and organelle distribution during cell division in vitro (alterations in the levels of small C terminal domain phosphatase-1 and -3, NuMa, and calnexin protein levels). This study sheds new light on the pathomechanisms of antidepressants action and their associated toxicity towards the reproductive system, emerging issues linked with animal or human reproductive health, and treatment of mood disorders.

Cereal grass juice in wound healing: hormesis and cell-survival in normal fibroblasts, in contrast to toxic events in cancer cells.

Natural products and traditional medicines are of great importance. Recent studies have demonstrated, that cereal grass juice improves wound healing, however the cellular and molecular mechanisms underlying these processes have not been fully characterized. Also, the full phytochemical characteristics of freshly squeezed juices obtained from cereal grasses is still missing. Thus, in this study a multi-dimensional analysis of juice parameters like refraction value, pH, chlorophyll and flavonoids content as well as antioxidant properties was performed. The results demonstrate that the effect induced by freshly squeezed cereal juices is strictly cell type-dependent. In this study, it is shown for the first time, that in normal fibroblasts (BJ cells) low dose cereal grass juices exhibit strong adaptive response through hormetic mechanism mediated by NF-κB/HO-1 and insulin/IGF-1 anti-oxidant pathways. As consequence, the process of wound healing is significantly upregulated. In cancer cells (ES-2 cells), despite anti-oxidant defense mechanism activation, levels of ROS and RNS are elevated. This leads to enhanced O-GlcNAcylation, DNA damage and cell cycle arrest, and as a result impaired wound healing. This study provides insights into the underlying mechanisms through which cereal grass juices activate hermetic adaptation response in normal fibroblasts, and induce cytotoxic and genotoxic events in cancer cells.

Melatonin receptors subtypes (MT1 and MT2) in the uterus masculinus of mature male european bison. Biological and seasonal reproductive role.

Regulation of seasonality in reproduction is closely related to melatonin and circadian rhythms. Melatonin affects the functions of reproductive organs through membrane melatonin receptors MT1 and MT2. The current knowledge about the presence, location and function of MT1 and MT2 receptors in the reproductive tract of an adult male European bison, seasonally breeding animal, is still missing. Frequently occurring organ in the male reproductive system of the European bison is uterus masculinus, what seems to confirm its specific role in seasonal reproduction control. Taking this into account, our study aimed to evaluate the expression of mRNA and protein synthesis for both melatonin receptors in the tissues of uterus masculinus in November and December in European bison. Protein synthesis of MT1 and MT2 receptors in uterus masculinus of mature European bisons was clearly raised in November and decreased in December. The comparable results were also found for mRNA expression of MT1 and MT2 receptors, where in November the expression level was significantly higher than in December. Therefore, we suggest that melatonin is needed in the European bison's reproductive system after a period of intensive reproductive activity in November. Probably, melatonin plays a protective function in uterus masculinus of the European bison and thus regulates the seasonal reproduction.

Local regulators of seasonal reproduction processes in uterus masculinus of an adult male European bison (Bison bonasus, Linnaeus 1758).

Growth factors, hypoxia-inducible factor 1-alpha (HIF-1α) and klotho protein all have very important functions in the male reproduction; however their role in the regulation of seasonal reproductive processes in the male European bison remains unclear. Similarly, although the uterus masculinus is very frequently found in the bison, its importance and functions remain unknown. It is likely that, this organ may have secretory functions and thus be a target for various regulatory factors. Therefore, the aim of this study was to investigate expression and activity of several factors: vascular endothelial growth factor (VEGF-A), fibroblast growth factor (FGF-2), transforming growth factor ß1 (TGF-ß1), nerve growth factor (NGF), insulin-like growth factor receptor (IGF-IR ß), hypoxia-inducible factor 1 alpha (HIF-1α), and klotho protein in the uterus masculinus, immediately after the season of the reproductive activity (November and December). Our study reveals that the growth factor expression levels are significantly higher in November, when compared to December, while expression of HIF-1α and klotho was higher in December. These results provide novel data on differences in the expression levels of several factors in the uterus maculinus of European bison bulls after the breeding season. The described factors may, therefore, be potent regulators of the seasonal reproduction.

Factor H uptake regulates intracellular C3 activation during apoptosis and decreases the inflammatory potential of nucleosomes.

Factor H (FH) binds apoptotic cells to limit the inflammatory potential of complement. Here we report that FH is actively internalized by apoptotic cells to enhance cathepsin L-mediated cleavage of endogenously expressed C3, which results in increased surface opsonization with iC3b. In addition, internalized FH forms complexes with nucleosomes, facilitates their phagocytosis by monocytes and induces an anti-inflammatory biased cytokine profile. A similar cytokine response was noted for apoptotic cells coated with FH, confirming that FH diminishes the immunogenic and inflammatory potential of autoantigens. These findings were supported by in vivo observations from CFH(-/-) MRL-lpr mice, which exhibited higher levels of circulating nucleosomes and necrotic cells than their CFH(+/+) littermates. This unconventional function of FH broadens the established view of apoptotic cell clearance and appears particularly important considering the strong associations with genetic FH alterations and diseases such as systemic lupus erythematosus and age-related macular degeneration.