Andreas Johannes Schmidt

Differential effects of glucocorticoids and gonadal steroids on glutathione levels in neuronal and glial cell systems

The aim of the present study was to investigate the short‐ and long‐term effects of glucocorticoids [corticosterone (CORT), dexamethasone (DEX), 6‐methylprednisolone (6‐MP)] and gonadal steroids [17β‐estradiol (E2), progesterone (PROG), testosterone (TEST)] on the levels of the antioxidant glutathione (GSH) in different cell systems of the CNS (neuronal hippocampal HT22 cells, primary hippocampal and neocortical brain cells, and C6 glioma cells). In HT22 cells, steroids exerted mainly long‐term effects. Significant increases of GSH levels were detectable after a 24 hr treatment with 10−7 M of DEX (122% ± 5%), 6‐MP (208% ± 32%), E2 (134% ± 10%), and TEST (155% ± 17%). A significant decrease occurred after incubation with PROG for 24 hr (79% ± 9%). In primary hippocampal cultures, a 24 hr treatment with DEX (140% ± 8%), E2 (123% ± 6%), and PROG (118% ± 5%) led to significant increases of the GSH levels, whereas, in neocortical primary cultures, only an incubation with E2 increased GSH (149% ± 8%). In C6 cells, hormone treatment led to both significant short‐term (1 hr: CORT 114% ± 5%, DEX 90% ± 3%, E2 88% ± 3%; 3 hr: DEX 115% ± 5%, E2 122% ± 6%, TEST 78% ± 4%) and significant long‐term (24 hr: CORT 74% ± 4%, 6‐MP 84% ± 5%, E2 115% ± 6%, PROG 91% ± 4%, TEST 116% ± 5%) effects. In summary, we were able to demonstrate differential effects of steroids on GSH levels in different cellular CNS models, showing an important influence of steroids and especially E2 on antioxidative cellular functions in neuronal and glial cells.

Effects of antidepressants on mRNA levels of antioxidant enzymes in human monocytic U-937 cells

Alterations of antioxidant enzyme activities have been described in a number of psychiatric disorders including major depression. Subsequently, the present study examined the effects of different types of antidepressants (desipramine, imipramine, maprotiline and mirtazapine) in different concentrations (10(-5), 10(-6) and 10(-7) M) on the mRNA levels of various enzymes of the antioxidant system, including both intracellular superoxide dismutase isoforms, glutathione peroxidase and catalase as well as several enzymes of the glutathione metabolism in monocytic U-937 cells after short- and long-term treatment (2.5 and 24 h) via RT-PCR. Results indicated mainly short-term decreases in the mRNA levels of antioxidant enzymes after treatment with these substances in all the concentrations used. In addition, after long-term treatment, significant increases in the mRNA levels were seen in the cases of Cu, Zn superoxide dismutase, gamma-glutamyl-cysteine synthetase, glutathione-S-transferase and glutathione reductase, including the impacts of all the antidepressants used in concentrations of 10(-6) M and 10(-7) M. Based on the large number of significant effects of all types of antidepressants tested on various antioxidant enzymes, we suggest that antioxidant enzymes may represent important targets in the course of antidepressive treatment.

Impact of haloperidol and quetiapine on the expression of genes encoding antioxidant enzymes in human neuroblastoma SH-SY5Y cells

Antipsychotics are known to alter antioxidant activities in vivo. Therefore, the aim of the present study was to examine in the human neuroblastoma SH-SY5Y cell line the impact of a typical (haloperidol) and an atypical (quetiapine) antipsychotic on the expression of genes encoding the key enzymes of the antioxidant metabolism (Cu, Zn superoxide dismutase; Mn superoxide dismutase; glutathione peroxidase; catalase) and enzymes of the glutathione metabolism (gamma-glutamyl cysteine synthetase, glutathione-S-transferase, gamma-glutamyltranspeptidase, glutathione reductase). The cells were incubated for 24h with 0.3, 3, 30 and 300microM haloperidol and quetiapine, respectively; mRNA levels were measured by polymerase chain reaction. In the present study, we observed mostly significant decreases of mRNA contents. With respect to the key pathways, we detected mainly effects on the mRNA levels of the hydrogen peroxide detoxifying enzymes. Among the enzymes of the glutathione metabolism, glutathione-S-transferase- and gamma-glutamyltranspeptidase-mRNA levels showed the most prominent effects. Taken together, our results demonstrate a significantly reduced expression of genes encoding for antioxidant enzymes after treatment with the antipsychotics, haloperidol and quetiapine.

Effects of steroid hormones on catalase activity in neuronal and glial cell systems

The aim of the present study was to determine short- (1 and 3 h) and long-term (24 h) effects of glucocorticoids [GCs; corticosterone (CORT), dexamethasone (DEX) and 6-methylprednisolone (6-MP)] and gonadal steroids [GSs; 17beta-estradiol (E2), progesterone (PROG) and testosterone (TEST)] on the activity of the hydrogen-peroxide-detoxifying enzyme catalase (CAT) in neural hippocampal HT22 cells and glial C6 cells because such effects have been described in peripheral organ systems. In HT22 cells, only long-term treatment with glucocorticoids (10(-5) M) induced effects on catalase activity, whereas gonadal steroids (10(-5) M) affected catalase activity after both short- and long-term incubations. At a lower concentration of 10(-7) M, glucocorticoids exerted only short-term treatment effects on catalase activity, while gonadal steroids (10(-7) M) affected the enzyme activity after short- and long-term treatments. In C6 glial cells, both glucocorticoids (10(-7) M) and gonadal steroids (10(-7) M) induced short- and long-term treatment effects. Thereby, our data show that steroid hormones differentially regulate catalase activity in models of the central nervous system (CNS) in a time- and steroid-dependent manner.

Antioxidative and steroid systems in neurological and psychiatric disorders

A large number of neurological and psychiatric diseases like Morbus Parkinson, amyotrophic lateral sclerosis, dementia, schizophrenia and probably also affective disorders show an enhanced production of reactive oxygen species. Moreover, alterations of antioxidative systems and beneficial effects of antioxidative substances including steroid compounds such as estrogens have been described in several of these diseases. This review focuses on alterations of antioxidative systems in the course of neurological diseases and psychiatric disorders and on the differential effects of steroids on these systems in the central nervous system. Moreover, a possible clinical relevance of alterations of circulating steroids and of steroid treatment under these conditions is discussed.

Effects of quetiapine, risperidone, 9-hydroxyrisperidone and ziprasidone on the survival of human neuronal and immune cells in vitro

Because there are reports on cytotoxic and cytoprotective effects of antipsychotics, the aim of the present study was to evaluate the impacts of different concentrations (1.6—50 μg/mL) of atypical antipsychotics on the survival of human neuronal (neuroblastoma SH-SY5Y) and immune (monocytic U-937) cells and on energy metabolism (ATP level after the incubation with antipsychotics in the concentration of 25 μg/mL). Statistical analysis showed that incubation for 24 h with the antipsychotics quetiapine, risperidone, 9-hydroxyrisperidone and ziprasidone led to a significantly enhanced cell survival in both cell lines in the lower concentrations. Higher concentrations exerted in part cytotoxic effects with the exception of quetiapine, but therapeutically relevant concentrations of the drugs were not cytotoxic in our experiments. Measurement of ATP contents in the neuronal cell line showed significantly increased levels after a 24-h treatment with 25 μg/mL risperidone and 9-hydroxyrisperidone. The other substances produced no effects. Our results show that the antipsychotic substances under investigation exert concentration-dependent effects on cell survival in both cell lines examined.

Impact of drugs approved for treating ADHD on the cell survival and energy metabolism: an in-vitro study in human neuronal and immune cells

The oxidative and antioxidative properties of psychostimulants such as methylphenidate and amphetamine are discussed controversially. The aim of the present study was to evaluate the impact of psychostimulants and atomoxetine in different concentrations between 31.25 and 5000 ng/ml on the survival of human neuronal (neuroblastoma SH-SY5Y) and immune (monocytic U-937) cells and the impact of psychostimulants and atomoxetine in different concentrations between 500 and 5000 ng/ml on energy metabolism (adenosine triphosphate [ATP] content) in SH-SY5Y cells. Statistical analysis revealed that incubation for 24 h with amphetamine led to a significantly enhanced cell survival in both cell lines after treatment with various (32.5, 125, 250 and 1250 ng/ml) concentrations. Methylphenidate and atomoxetine induced a significantly enhanced cell survival at lower concentrations in the SH-SY5Y cell line, whereas in the U-937 cell line higher concentrations increased the cell survival. Incubation with the highest concentration of methylphenidate (5000 ng/ml) caused a significant reduction of cell survival in both cell types. Measurement of ATP contents in the neuronal cell line revealed no significant effects of the investigated compounds. Our results show that the examined substances exert concentration-dependent effects on cell survival in both applied cell lines.

Interleukin-6 induces glutathione in hippocampal cells.

The cytokine interleukin-6 (IL-6) increases the levels of the physiological antioxidant glutathione (GSH) in peripheral organ systems such as liver tissue. Only little evidence exists about the actions of this cytokine on GSH in neuronal cell systems despite its possible neuroprotective effects. Therefore, we here characterized the effects of IL-6 on GSH in clonal hippocampal HT22 cells and in rat neuronal primary hippocampal cells. Our results demonstrate significant increases of GSH under most conditions after treatment with IL-6 in a time range of 1 to 48 h (HT22 cells) and 1 to 72 h (primary rat neuronal hippocampal cells). Further studies with an IL-6 antibody strongly support the specificity of the effects. These results suggest that IL-6 plays a substantial role in the regulation of GSH in hippocampal cells.

Impact of Plant Extracts Tested in Attention-Deficit/Hyperactivity Disorder Treatment on Cell Survival and Energy Metabolism in Human Neuroblastoma SH-SY5Y Cells

Plant extracts such as Hypericum perforatum and Pycnogenol® have been tested as alternatives to the classical ADHD drugs. It has been possible to describe neuroprotective effects of such plant extracts. A reduction of ADHD symptoms could be shown in clinical studies after the application of Pycnogenol®, which is a pine bark extract. The impacts of the standardized herbal extracts Hypericum perforatum, Pycnogenol® and Enzogenol® up to a concentration of 5000 ng/mL on cell survival and energy metabolism in human SH‐SY5Y neuroblastoma cells has been investigated in the present examination. Hypericum perforatum significantly decreased the survival of cells after treatment with a concentration of 5000 ng/mL, whereas lower concentrations exerted no significant effects. Pycnogenol® induced a significant increase of cell survival after incubation with a concentration of 32.25 ng/mL and a concentration of 250 ng/mL. Other applied concentrations of Pycnogenol® failed to exert significant effects. Treatment with Enzogenol® did not lead to significant changes in cell survival.

Hydrogen sulfide affects radical formation in the hippocampus of LPS treated rats and the effect of antipsychotics on hydrogen sulfide forming enzymes in human cell lines.

Objectives: Psychiatric disorders, such as schizophrenia and other neuroinflammatory diseases are accompanied by an increase in the oxidative stress and changes in the immune system and in the metabolic, hormonal and neurological components of the central nervous system (CNS). Hydrogen sulfide (H2S) is a gaseous molecule that is endogenously produced in the peripheral and central nervous system through cysteine by the following major H2S producing enzymes in the brain: cystathionine-γlyase (CSE), cystathionine ß-synthase (CBS) and 3-mercaptopyruvate sulfurtransferase (MPST). The physiological effects of H2S are broad, with antioxidative properties being a major role in the body. The aims of our investigation were to analyze the central nervous antioxidant, metabolic and neuronal effects in the hippocampus of the rat after inflammatory peripheral lipopolysaccharide (LPS) treatment; and to examine the effects of antipsychotics on the expression of these enzymes in human cell lines. Material and Methods: Male Lewis rats (250 g) received an i.p. LPS injection (1 mg/kg) 24 h before microdialysis experiments. Conscious rats were infused via these probes (1.5 μl/min) with a radical scavenger 1-hydroxy-3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine (CMH) in Krebs-Ringer solution. Sodiumhydrogensulfide (NaHS, 10 μg/min) was infused after a 2- h baseline for 1 h. Corticosterone, glutamate, glucose and lactate were measured by Elisa. Reactive oxygen species (ROS) were detected by electron spin resonance spectroscopy (ESR). The impact of the antipsychotics haloperidol, clozapine, olanzapine and risperidone on the expression of genes encoding the key enzymes of H2S synthesis was studied at the human neuroblastoma SH-SY5Y and monocytic U-937 cell lines. The cells were incubated for 24 h with 30 μM antipsychotic following which mRNA levels were measured by polymerase chain reaction. Results: Microdialysate glucose and lactate levels dramatically increased in the hippocampus of LPS untreated rats by local application of NaHS. By contrast, in the LPS pretreated rats, there was no effect of NaHS infusion on glucose but a further significant increase in microdialysate lactate was found. It was LPS pretreatment alone that particularly enhanced lactate levels. There was a marked increase in hippocampal microdialysate glutamate levels after local NaHS infusion in LPS untreated animals. In LPS treated rats, no change was observed by NaHS, but LPS itself had the strongest effect on microdialysate glutamate levels. Microdialysate corticosterone levels were reduced by NaHS in both LPS pretreated and untreated rats. The formation of free radicals in the hippocampus significantly reduced in LPS pretreated rats, while in LPS untreated rats a significant increase was observed after NaHS infusion. In human SH-SY5Y and U-937 cells, all three major enzymes of H2S-Synthesis, namely cystathionine-γ-lyase, cystathione ß-synthase and 3-mercaptopyruvate sulfurtransferase, could be detected by PCR. The antipsychotics haloperidol, clozapine, olanzapine and risperidone affected all three enzymes in different ways; with haloperidol and risperidone showing major effects that led to reductions in CBS or CSE expression. Discussion: The local application of NaHS in the hippocampus of the rat strongly affected glucose, lactate and glutamate release. Contrastingly, in LPS pretreated rats, a decreased radical formation was the only effect found. H2S synthetizing enzymes may be involved in antipsychotic mechanisms, although no clear common mechanism could be found.