The role of vitamin D₃ and its receptor in the female reproductive system

Badania ostatnich lat sugerują pogłębiający się problem niedoboru witaminy D3. Oprócz dobrze poznanej roli witaminy D3 w utrzymaniu homeostazy wapniowo-fosforanowej, coraz większym zainteresowaniem cieszy się jej udział w regulacji funkcji żeńskiego układu rozrodczego. Występowanie receptora witaminy D3 w jajnikach, endometrium macicy oraz łożysku świadczy, że są to tkanki docelowe dla tej witaminy. Liczne badania potwierdzają, iż obniżony poziom aktywnych form witaminy D3 sprzyja zaburzeniom budowy i funkcji jajnika oraz macicy. Dotychczas zaobserwowano istotną rolę kalcytriolu w procesach folikulogenezy, steroidogenezy, implantacji zarodka, przebiegu ciąży oraz utrzymaniu zdrowia potomstwa. Obniżony poziom witaminy D3 sprzyja patologiom żeńskiego układu rozrodczego, tj. zespół policystycznych jajników, przedwczesna niewydolność jajników, endometrioza czy mięśniaki macicy. Ekspozycja na promieniowanie słoneczne i zbilansowana dieta, pozwalają na utrzymanie prawidłowego stężenia kalcydiolu w surowicy krwi. Poszerzenie wiedzy na temat roli witaminy D3 w patogenezie schorzeń żeńskiego układu rozrodczego, może przyczynić się do wykorzystania jej suplementacji jako skutecznego sposobu profilaktyki i leczenia.Recent studies suggested an increasing problem with deficiency of vitamin D3. Except for the well-known role of vitamin D3 in the maintenance of calcium and phosphate homeostasis, its contribution to the regulation of the female reproduction becomes more popular. The presence of vitamin D3 receptor in the ovary, endometrium and placenta indicates that these tissues are target for vitamin D3 action. Many studies confirm that low level of active vitamin D3 promotes the occurrence of abnormalities in the structure and function of the ovary and uterus. To date, a significant role of calcitriol in folliculogenesis, steroidogenesis, embryo implantation, pregnancy and the offspring health has been observed. Decreased level of vitamin D3 associated with many pathologies in the female reproductive system such as polycystic ovarian syndrome, premature ovarian failure, endometriosis or uterine fibroids. Exposure to sunlight and balanced diet help to maintain normal levels of calcidiol in blood serum. Increasing knowledge about the role of vitamin D3 in female reproductive pathogenesis may contribute to the use of adequate vitamin D3 supplementation as an effective tool in the preventive and therapeutic treatment

Bioanalytical methodology for determination of glutamate and aspartate for use in pharmacological sciences with application of Integrated Calibration Method

A new calibration approach based on the adaptation of Integrated Calibration Method (ICM) to consecutive two components analysis in separation science is presented. Consecutive ICM method (C-ICM) was conceptually developed and applied to determination of two excitatory amino acids - glutamate and aspartate in cerebrospinal fluids collected by the use of brain microdialysis from freely-moving animals. Both analytes as a neurotransmitters play an important role in formation of the memory trace, and thus the processes of learning and memory. Due to their low concentration and presence of interferences, considered analytical system - animal brain - was a big challenge. High-performance liquid chromatography (HPLC) with electrochemical detection (ECD) was used in all experimental work. The most important feature of proposed method is integration of interpolative and extrapolative ways to calculate analyte concentration in single calibration procedure, which consequently leads to obtain series of six estimations of analytical result. Comparison of individual estimations with each other allows for a more in-depth analysis of systematic errors. It was proved that C-ICM approach enables diagnosis and compensation of systematic errors induced by occurrence of interference effects and improvement of accuracy of analytical results. Most of all, it was demonstrated that application of this method is efficient and useful analytical tool in analysis of complicated biological samples in pharmacology and neuroscience

Gut Inflammation Induced by Finasteride Withdrawal: Therapeutic Effect of Allopregnanolone in Adult Male Rats

The treatment with finasteride (i.e., an inhibitor of 5α-reductase) may be associated with different side effects (i.e., depression, anxiety, cognitive impairment and sexual dysfunction) inducing the so-called post finasteride syndrome (PFS). Moreover, previous observations in PFS patients and an experimental model showed alterations in gut microbiota populations, suggesting an inflammatory environment. To confirm this hypothesis, we have explored the effect of chronic treatment with finasteride (i.e., for 20 days) and its withdrawal (i.e., for 1 month) on the levels of steroids, neurotransmitters, pro-inflammatory cytokines and gut permeability markers in the colon of adult male rat. The obtained data demonstrate that the levels of allopregnanolone (ALLO) decreased after finasteride treatment and after its withdrawal. Following the drug suspension, the decrease in ALLO levels correlates with an increase in IL-1β and TNF-α, serotonin and a decrease in dopamine. Importantly, ALLO treatment is able to counteract some of these alterations. The relation between ALLO and GABA-A receptors and/or pregnenolone (ALLO precursor) could be crucial in their mode of action. These observations provide an important background to explore further the protective effect of ALLO in the PFS experimental model and the possibility of its translation into clinical therapy

Neurotoxicological profile of the hallucinogenic compound 25I-NBOMe

4-Iodo-2,5-dimethoxy-N-(2-methoxybenzyl)phenethylamine (25I-NBOMe) is a new psychoactive substance with strong hallucinogenic properties. Our previous data reported increased release of dopamine, serotonin, and glutamate after acute injections and a tolerance development in the neurotransmitters release and rats’ behavior after chronic treatment with 25I-NBOMe. The recreational use of 25I-NBOMe is associated with severe intoxication and deaths in humans. There is no data about 25I-NBOMe in vivo toxicity towards the brain tissue. In this article 25I-NBOMe-crossing through the blood–brain barrier (BBB), the impact on DNA damage, apoptosis induction, and changes in the number of cortical and hippocampal cells were studied. The presence of 25I-NBOMe in several brain regions shortly after the drug administration and its accumulation after multiple injections was found. The DNA damage was detected 72 h after the chronic treatment. On the contrary, at the same time point apoptotic signal was not identified. A decrease in the number of glial but not in neural cells in the frontal (FC) and medial prefrontal cortex (mPFC) was observed. The obtained data indicate that 25I-NBOMe passes easily across the BBB and accumulates in the brain tissue. Observed oxidative DNA damage may lead to the glial cells’ death

Effect of Psilocybin and Ketamine on Brain Neurotransmitters, Glutamate Receptors, DNA and Rat Behavior

Clinical studies provide evidence that ketamine and psilocybin could be used as fast-acting antidepressants, though their mechanisms and toxicity are still not fully understood. To address this issue, we have examined the effect of a single administration of ketamine and psilocybin on the extracellular levels of neurotransmitters in the rat frontal cortex and reticular nucleus of the thalamus using microdialysis. The genotoxic effect and density of glutamate receptor proteins was measured with comet assay and Western blot, respectively. An open field test, light–dark box test and forced swim test were conducted to examine rat behavior 24 h after drug administration. Ketamine (10 mg/kg) and psilocybin (2 and 10 mg/kg) increased dopamine, serotonin, glutamate and GABA extracellular levels in the frontal cortex, while psilocybin also increased GABA in the reticular nucleus of the thalamus. Oxidative DNA damage due to psilocybin was observed in the frontal cortex and from both drugs in the hippocampus. NR2A subunit levels were increased after psilocybin (10 mg/kg). Behavioral tests showed no antidepressant or anxiolytic effects, and only ketamine suppressed rat locomotor activity. The observed changes in neurotransmission might lead to genotoxicity and increased NR2A levels, while not markedly affecting animal behavior

Diabetic Encephalopathy in a Preclinical Experimental Model of Type 1 Diabetes Mellitus: Observations in Adult Female Rat

Patients affected by diabetes mellitus (DM) show diabetic encephalopathy with an increased risk of cognitive deficits, dementia and Alzheimer’s disease, but the mechanisms are not fully explored. In the male animal models of DM, the development of cognitive impairment seems to be the result of the concomitance of different processes such as neuroinflammation, oxidative stress, mitochondrial dysfunction, and aberrant synaptogenesis. However, even if diabetic encephalopathy shows some sex-dimorphic features, no observations in female rats have been so far reported on these aspects. Therefore, in an experimental model of type 1 DM (T1DM), we explored the impact of one month of pathology on memory abilities by the novel object recognition test and on neuroinflammation, synaptogenesis and mitochondrial functionality. Moreover, given that steroids are involved in memory and learning, we also analysed their levels and receptors. We reported that memory dysfunction can be associated with different features in the female hippocampus and cerebral cortex. Indeed, in the hippocampus, we observed aberrant synaptogenesis and neuroinflammation but not mitochondrial dysfunction and oxidative stress, possibly due to the results of locally increased levels of progesterone metabolites (i.e., dihydroprogesterone and allopregnanolone). These observations suggest specific brain-area effects of T1DM since different alterations are observed in the cerebral cortex