Fedotcheva T.A.,Russian National Research Medical University |
Shirokikh K.E.,Russian National Research Medical University |
Matyushin A.I.,Russian National Research Medical University |
Rzheznikov V.M.,Institute of Experimental Endocrinology |
And 2 more authors.
Biophysics (Russian Federation) | Year: 2015
In order to determine the cytotoxic or cytoprotective effect of the synthetic isoflavonoid genistein, we studied its effect on HeLa tumor cells, which contain estrogen alpha receptors and do not contain estrogen beta receptors. It was shown that the genistein concentration (IC50 = 0.2 mM) at which the half maximal inhibition of the HeLa cell viability is achieved is ten times higher than the concentrations of tamoxifen and cisplatin, which are reference agents with a cytotoxic effect. At micromolar concentrations (0.1–10 µM) genistein decreased the cytotoxic effects of cisplatin and tamoxifen. We found the reduced Bax mRNA expression and increased Bcl-2 mRNA expression during incubation of the cells with genistein, which also indicates its cytoprotective anti-apoptotic effect. Genistein, even in high concentrations, had no effect on the membrane potential and calcium capacity of isolated mitochondria and did not activate the opening of the Ca2+-induced mitochondrial pore. Thus, the data show a protective effect of the isoflavonoid genistein on tumor cells. © 2015, Pleiades Publishing, Inc.
Sparks L.M.,Pennington Biomedical Research Center |
Sparks L.M.,Maastricht University |
Moro C.,Pennington Biomedical Research Center |
Moro C.,University Paul Sabatier |
And 8 more authors.
PLoS ONE | Year: 2011
Objective: Disturbances in lipid metabolism are strongly associated with insulin resistance and type 2 diabetes (T2D). We hypothesized that activation of cAMP/PKA and calcium signaling pathways in cultured human myotubes would provide further insight into regulation of lipid storage, lipolysis, lipid oxidation and insulin responsiveness. Methods: Human myoblasts were isolated from vastus lateralis, purified, cultured and differentiated into myotubes. All cells were incubated with palmitate during differentiation. Treatment cells were pulsed 1 hour each day with forskolin and ionomycin (PFI) during the final 3 days of differentiation to activate the cAMP/PKA and calcium signaling pathways. Control cells were not pulsed (control). Mitochondrial content, 14C lipid oxidation and storage were measured, as well as lipolysis and insulin-stimulated glycogen storage. Myotubes were stained for lipids and gene expression measured. Results: PFI increased oxidation of oleate and palmitate to CO2 (p<0.001), isoproterenol-stimulated lipolysis (p = 0.01), triacylglycerol (TAG) storage (p<0.05) and mitochondrial DNA copy number (p = 0.01) and related enzyme activities. Candidate gene and microarray analysis revealed increased expression of genes involved in lipolysis, TAG synthesis and mitochondrial biogenesis. PFI increased the organization of lipid droplets along the myofibrillar apparatus. These changes in lipid metabolism were associated with an increase in insulin-mediated glycogen storage (p<0.001). Conclusions: Activation of cAMP/PKA and calcium signaling pathways in myotubes induces a remodeling of lipid droplets and functional changes in lipid metabolism. These results provide a novel pharmacological approach to promote lipid metabolism and improve insulin responsiveness in myotubes, which may be of therapeutic importance for obesity and type 2 diabetes. © 2011 Sparks et al.
Laukova M.,Institute of Experimental Endocrinology |
Vargovic P.,Institute of Experimental Endocrinology |
Csaderova L.,Slovak Academy of Sciences |
Chovanova L.,Institute of Experimental Endocrinology |
And 4 more authors.
NeuroImmunoModulation | Year: 2012
Objectives: Stress-induced rise in circulating catecholamines (CAs), followed by modulation of β-adrenergic receptors (adrenoceptors, ARs), is one of the pathways involved in the stress-mediated effects of immune functions. The spleen is an organ with a high number of lymphocytes and provides a unique microenvironment in which they reside. Thus, lymphocytes may respond differently to CAs in the spleen than in the circulation. No reports exist concerning the involvement of β-ARs in stress-mediated effects on T and B cells isolated from the spleen. Therefore, our aim was to investigate the effect of single stress exposure on gene expression and cellular localization of β-adrenoceptor subtypes in splenic T and B cells. We tried to correlate changes in adrenoceptors with the expression of apoptotic proteins. Methods: Immobilization (IMMO) was used as a stress model. T and B cells were isolated from rat spleen using magnetically labeled antibodies. The gene expression of individual adrenoceptors and apoptotic proteins was evaluated by real-time PCR. Immunofluorescence was used to evaluate localization and adrenoceptor expression. Results: We have found T cells to be more vulnerable to stress compared to B cells, because of increased β 1-, β 2-and β 3-ARs after a single IMMO. Moreover, β 2-ARs translocated from the nucleus to the plasma membrane in T cells after IMMO. The rise in β-ARs most probably led to the rise of Bax mRNA and Bax to Bcl-2 mRNA ratio. This might suggest the induction of an apoptotic process in T cells. Conclusion: Higher susceptibility of T cells to stress via modulation of β-ARs and apoptotic proteins might shift the immune responsiveness in the spleen. Copyright © 2012 S. Karger AG, Basel.
Smirnova Z.S.,Blokhin Oncological Research Center |
Rzheznikov V.M.,Institute of Experimental Endocrinology |
Tolkachev V.N.,Blokhin Oncological Research Center |
Borisova L.M.,Blokhin Oncological Research Center |
And 6 more authors.
Eksperimental'naya i Klinicheskaya Farmakologiya | Year: 2014
Cytestrole acetate (CA), in the structure of which the steroidal antiestrogen component is associated with bis-β-cloroethylamino group, exhibits a strong cytotoxic activity against hormone-dependent cancer cell lines (CaOV, HeLa, MCF-7). In doxorubicin-resistant MCF-7 cells, CA potentiates the cytotoxic effect of etoposide and doxorubicin, and the IC50 for CA in these cells is 40 times lower than that for tamoxifen (TAM). In transplantable mice breast adenocarcinoma Ca-755, the therapeutic CA dose is 25 mg/kg when administered subcutaneously in oil solution for 5 days. On the DMBA-induced mammary tumors in rats, CA injected subcutaneously led to partial regressions 4 weeks after treatment in 75% of test rats, whereas TAM produced this effect in 43% of rats. Among various drug forms of CA, the most active were oil solution of CA in gelatin capsules for oral use and liposomal emulsion for intravenous administration, since these forms exhibited the highest values of Ca-755 tumor growth inhibition index (TGI = 97-98%).
Zatkova M.,Comenius University |
Bakos J.,Comenius University |
Bakos J.,Institute of Experimental Endocrinology |
Hodosy J.,Comenius University |
And 2 more authors.
Biomedical Papers | Year: 2016
Background. Recent research has produced an explosion of experimental data on the complex neurobiological mechanisms of developmental disorders including autism. Animal models are one approach to studying the phenotypic features and molecular basis of autism. In this review, we describe progress in understanding synaptogenesis and alterations to this process with special emphasis on the cell adhesion molecules and scaffolding proteins implicated in autism. Genetic mouse model experiments are discussed in relation to alterations to selected synaptic proteins and consequent behavioral deficits measured in animal experiments. Methods. Pubmed databases were used to search for original and review articles on animal and human clinical studies on autism. Results. The cell adhesion molecules, neurexin, neurolignin and the Shank family of proteins are important molecular targets associated with autism. Conclusion. The heterogeneity of the autism spectrum of disorders limits interpretation of information acquired from any single animal model or animal test. We showed synapse-specific/ model-specific defects associated with a given genotype in these models. Characterization of mouse models with genetic variations may help study the mechanisms of autism in humans. However, it will be necessary to apply new analytic paradigms in using genetically modified mice for understanding autism etiology in humans. Further studies are needed to create animal models with mutations that match the molecular and neural bases of autism. © 2016, PALACKY UNIV. All rights reserved.
Spolcova A.,Czech Institute of Organic Chemistry And Biochemistry |
Mikulaskova B.,Czech Institute of Organic Chemistry And Biochemistry |
Krskova K.,Institute of Experimental Endocrinology |
Gajdosechova L.,Institute of Experimental Endocrinology |
And 6 more authors.
BMC Neuroscience | Year: 2014
Background: Insulin signaling and Tau protein phosphorylation in the hippocampi of young and old obese Zucker fa/fa rats and their lean controls were assessed to determine whether obesity-induced peripheral insulin resistance and aging are risk factors for central insulin resistance and whether central insulin resistance is related to the pathologic phosphorylation of the Tau protein. Results: Aging and obesity significantly attenuated the phosphorylation of the insulin cascade kinases Akt (protein kinase B, PKB) and GSK-3β (glycogen synthase kinase 3β) in the hippocampi of the fa/fa rats. Furthermore, the hyperphosphorylation of Tau Ser396 alone and both Tau Ser396 and Thr231 was significantly augmented by aging and obesity, respectively, in the hippocampi of these rats. Conclusions: Both age-induced and obesity-induced peripheral insulin resistance are associated with central insulin resistance that is linked to hyperTau phosphorylation. Peripheral hyperinsulinemia, rather than hyperglycemia, appears to promote central insulin resistance and the Tau pathology in fa/fa rats. © 2014 Špolcová et al.; licensee BioMed Central Ltd.
Mul J.D.,University Utrecht |
Mul J.D.,University of Cincinnati |
O'Duibhir E.,University Utrecht |
Shrestha Y.B.,Georgia State University |
And 8 more authors.
PLoS ONE | Year: 2013
The orexigenic neuropeptide melanin-concentrating hormone (MCH), a product of Pmch, is an important mediator of energy homeostasis. Pmch-deficient rodents are lean and smaller, characterized by lower food intake, body-, and fat mass. Pmch is expressed in hypothalamic neurons that ultimately are components in the sympathetic nervous system (SNS) drive to white and interscapular brown adipose tissue (WAT, iBAT, respectively). MCH binds to MCH receptor 1 (MCH1R), which is present on adipocytes. Currently it is unknown if Pmch-ablation changes adipocyte differentiation or sympathetic adipose drive. Using Pmch-deficient and wild-type rats on a standard low-fat diet, we analyzed dorsal subcutaneous and perirenal WAT mass and adipocyte morphology (size and number) throughout development, and indices of sympathetic activation in WAT and iBAT during adulthood. Moreover, using an in vitro approach we investigated the ability of MCH to modulate 3T3-L1 adipocyte differentiation. Pmch-deficiency decreased dorsal subcutaneous and perirenal WAT mass by reducing adipocyte size, but not number. In line with this, in vitro 3T3-L1 adipocyte differentiation was unaffected by MCH. Finally, adult Pmch-deficient rats had lower norepinephrine turnover (an index of sympathetic adipose drive) in WAT and iBAT than wild-type rats. Collectively, our data indicate that MCH/MCH1R-pathway does not modify adipocyte differentiation, whereas Pmch-deficiency in laboratory rats lowers adiposity throughout development and sympathetic adipose drive during adulthood. © 2013 Mul et al.
Bohnekamp J.,University of Leipzig |
Boselt I.,University of Leipzig |
Saalbach A.,University of Leipzig |
Tonjes A.,University of Leipzig |
And 11 more authors.
Biochemical and Biophysical Research Communications | Year: 2010
Chemokine receptors control leukocyte chemotaxis and cell-cell communication but have also been associated with pathogen entry. GPR33, an orphan member of the chemokine-like receptor family, is a pseudogene in most humans. After the appearance of GPR33 in first mammalian genomes, this receptor underwent independent pseudogenization in humans, other hominoids and some rodent species. It was speculated that a likely cause of GPR33 inactivation was its interplay with a rodent-hominoid-specific pathogen. Simultaneous pseudogenization in several unrelated species within the last 1 million years (myr) caused by neutral drift appears to be very unlikely suggesting selection on the GPR33 null-allele. Although there are no signatures of recent selection on human GPR33 we found a significant increase in the pseudogene allele frequency in European populations when compared with African and Asian populations. Because its role in the immune system was still hypothetical expression analysis revealed that GPR33 is highly expressed in dendritic cells (DC). Murine GPR33 expression is regulated by the activity of toll-like receptors (TLR) and AP-1/NF-κB signaling pathways in cell culture and in vivo. Our data indicate an important role of GPR33 function in innate immunity which became dispensable during human evolution most likely due to past or balancing selection. © 2010 Elsevier Inc. All rights reserved.
Tacke M.,Applied Cachexia Research |
Ebner N.,Applied Cachexia Research |
Boschmann M.,Experimental and Clinical Research Center |
Jarius A.,Experimental and Clinical Research Center |
And 10 more authors.
Journal of the American Medical Directors Association | Year: 2013
Objectives: Muscle wasting is common in patients with chronic heart failure (HF) and worsens functional status. Protein catabolism is characteristic of muscle wasting and contributes to resting energy expenditure (REE). Glucagonlike peptide 1 (GLP-1) is linked to REE in healthy individuals. We aimed to evaluate (1) whether REE is elevated in patients with HF with muscle wasting, and (2) whether basal GLP-1 levels are linked to REE in HF. Design: Cross-sectional study. Setting: Ambulatory patients with HF were recruited at the Charité Medical School, Campus Virchow-Klinikum, Berlin, Germany. Participants: A total of 166 patients with HF and 27 healthy controls participating in the Studies Investigating Co-morbidities Aggravating Heart Failure (SICA-HF) were enrolled. GLP-1 was measured in 55 of these patients. Measurements: Body composition was measured by dual-energy X-ray absorptiometry (DEXA). Muscle wasting was defined as appendicular lean mass of at least 2 SDs below values of a healthy young reference group. REE was measured by indirect calorimetry. GLP-1 was assessed by ELISA. Results: Thirty-four of 166 patients (mean age 67.4 ± 10.2 years, 77.7% male, New York Heart Association class 2.3 ± 0.6) presented with muscle wasting. REE in controls and patients with muscle wasting was significantly lower than in patients without muscle wasting (1579 ± 289 and 1532 ± 265 vs 1748 ± 359 kcal/d, P= .018 and P= .001, respectively). REE normalized for fat-free mass (FFM) using the ratio method (REE/FFM) and analysis of covariance was not different (P= .23 and .71, respectively). GLP-1 did not significantly correlate with REE (P= .49), even not after controlling for FFM using multivariable regression (P= .15). Conclusions: Differences in REE are attributable to lower FFM. GLP-1 does not relate to REE in patients with HF, possibly because of HF-related effects on REE. © 2013 American Medical Directors Association, Inc.