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1

The influence of IL-10 on the inflammatory reaction changes in the mice after 1-methyl-4-phenyl-1,2,3,6-tertahydropyridine (MPTP) treatment

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Sznejder A., Joniec-Maciejak I., Ciesielska A., Schwenkgrub J., Wawer A., Bankiewicz K., Czlonkowska A., Czlonkowski A.
Acta Neurobiologiae Experimentalis
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2011
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tom 71
|
nr S
Parkinson’s disease (PD) is one of most frequent neurological disorder characterized by the loss of dopaminergic neurons in substantia nigra and striatum. The typical reaction of central neural system (CNS) on neurodegenerative processes is microglia activation and the inflammatory reaction. Microglia activation stimulates astrocytes response, playing important role in neuroimmune reaction. Microglia cells secrete two types of mediators of the inflammatory process: anti- and pro- inflammatory. In the first stage of Parkinson’s disease, pro-inflammatory cytokines have important meaning. We investigated the effect of an adenoassociated viral vector (AAV2) containing the complementary DNA (cDNA) for human interleukine 10 (hIL-10). The aim of the present study was to examine the evaluation of inflammatory reaction changes following increased concentration of hIL-10 in the murine model of PD induced by MPTP. Male C57BL mice 12 month-old were used in this study. AAV2 vector was bilateraly administered into striatum at 7, 21, 28 days prior to MPTP intoxication. We observed changes in the morphology of microglia cells, infiltration of lymphocytes T (population of CD3+, CD4+ and CD8+) and some differences in the level of one of the most important pro inflammatory cytokines – IL-1α. Our study showed that IL-10 is strongly involved in the inflammatory reaction in the murine model of Parkinson’s disease induced by MPTP. After MPTP intoxication we observed the increase of activated microglia cells, infiltration of lymphocytes T and higher level of IL-1α mRNA. AAV2-hIL-10-treated mice displayed a significant decrease in the activated microglia cells, elevated expression of IL-10 receptors observed on glia cells, strong infiltration of lymphocytes T (mainly CD4+ and CD3+, less CD8+) and minor expression of IL-1α mRNA. Further research must be conducted to provide more evidence of protective role of IL-10 in Parkinson disease.
2

The effect of human interleukin-10 on the nitric oxide synthases expression in MPTP- based model of Parkinson's disease

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Schwenkgrub J., Joniec-Maciejak I., Ciesielska A., Sznejder A., Wawer A., Bankiewicz K., Czlonkowska A., Czlonkowski A.
Acta Neurobiologiae Experimentalis
|
2011
|
tom 71
|
nr S
Parkinson’s disease (PD) is a progressive degenerative disorder, which etiology and pathogenesis remains unknown. Post mortem analysis of PD brain and studies on neurotoxic animal models of PD have provided evidence to support the involvement of oxidative stress and neuroinflammatory processes in the pathogenesis of PD. The high level of nitric oxide (NO) is produced by iNOS during the neuroinflammatory process caused by MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) treatment. Under pathological condition NO can easily react with superoxide to form peroxynitrite (ONOOˉ), which is a strong oxidant. In the present study was examined the influence of the increased concentration of IL-10 (an anti-inflammatory cytokine) on the NOS expression in mouse model of PD induced by MPTP. One year-old male C57Bl mice were used in this study. An adeno-associated viral vector expressing the gene for human interleukin-10 (hIL-10) was used to transduce striatal cells 4 weeks prior to MPTP intoxication. Mice were sacrificed at the different time intervals: 1, 7 and 21 days after MPTP injection. Immunohistochemical and western blot analyses provide evidence for the protective properties of AAV2-hIL-10 in the MPTP-induced model of PD. There were reduction in the dopaminergic neuron quantity in SNpc and tyrosine hydroxylase protein in the striatum after MPTP injections, whereas in the group additionally treated with AAV2-hIL10 neuroprotection was observed. Treatment with AAV2-hIL-10 suppressed the MPTP-induced increase in iNOS and 3-nitrotyrosine (3-NT) expression in the midbrain.
3

Effect of AAF-2-h1L-10 on immune response following toxic degeneration caused by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine in micep.345

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Joniec L., Ciesielska A., Gladka A., Schwenkgrub J., Sznejder A., Cudna A., Hadaczek P., Bankiewicz K., Czlonkowska A., Czlonkowski A.
Acta Neurobiologiae Experimentalis
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2009
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tom 69
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nr 3
Interleukine 10 (IL-10) – an antiinfl ammatory cytokine produced by lymphocytes and mononuclear phagocytes including microglia. IL10 modulates the biological activities of immune cells resulting in a decreased production of pro-infl ammatory mediators including cytokines, chemokines and adhesion molecules. The aim of the present study was to examine the effect of an adeno-associated viral type-2 (AAV2) vector containing human interleukin-10 (hIL-10) gene to evaluation of immune response to the AAV2-hIL-10 (measured as IFN-γ, GFAP and TGFβ mRNA expression) in the murine model of Parkinson’s disease (PD) induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Male C57BL/6 mice 12 months-old were used in this study. MPTP (40 mg/kg) was injected in four intraperitoneal injections at 1-h intervals. AAV2-hIL-10 vector was bilaterally administered into striatum at 14, 21 or 28 days prior to MPTP intoxication. Animals were sacrifi ced by spinal cords dislocation at 7 days following MPTP injection. TGFβ, IFN-γ and GFAP mRNA expression was examined by RT-PCR method. MPTP treatment signifi cantly increased IFN-γ mRNA expression. AAV2-hIL-10 administration strongly increased IFN-γ as well as TGFβ and GFAP (21 and 28 day) gene expression compared to control and MPTP group. Our results point to the necessity of the reinterpretation of the role of the infl ammatory reaction in nerodegenerative processes
4

The influence of AAV-mediated gene transfer of human interleukin 10 on the neurodegenerative process in the murine model of parkinson's disease

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Wawer A., Joniec-Maciejak I., Schwenkgrub J., Sznejder A., Ciesielska A., Bankiewicz K., Czlonkowska A., Czlonkowski A.
Acta Neurobiologiae Experimentalis
|
2011
|
tom 71
|
nr S
Parkinson’s disease (PD) is one of the most common neurodegenerative disorders. It is characterized by a progressive loss of dopaminergic neurons, which occurs mainly in the substantia nigra (SN), resulting in the loss of nerve terminals, accompanied by a decreased concentration of dopamine (DA) and its metabolites in the striatum. Concurrently with neurodegeneration, a chronic inflammation occurs in the affected regions of the brain. Mechanisms and etiology of the neurodegeneration are still unknown. One potential strategy for therapy of PD is to reduce the neurodegeneration by inhibiting the inflammatory reaction. Nowadays, there are high hopes for the gene therapy based treatment. This involves using a noninfectious virus administered directly into the brain. In this study a transfer of AAV vector containing the complementary DNA for human interleukin 10 (IL-10) into the nigrostriatal pathway was used. IL-10 is one of the major antiinflammatory cytokines. The aim of the present study was to examine the expression of human IL-10, administered into striatum by viral vector AAV2-hIL-10 and investigate the influence of this cytokine on neurodegenerative process in the murine model of PD induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). One year old male C57Bl mice were used in this study. The expression of human IL-10 was measured by enzyme-linked immunosorbent assay (ELISA). To evaluate the influence of the human IL-10 on neurodegeneration concentrations of striatal DA, 3,4-dihydroxyphenyl acetic acid and homovanillic acid were measured by high performance liquid chromatography (HPLC). The findings demonstrate human IL-10 secretion in the mouse brain after striatal infusion of AAV2-hIL-10. This study suggests that human IL-10 delivered by an AAV2 vector preserves nigrostriatal function after MPTP intoxication (lower decrease in DA concentration). IL-10 can play a vital role in inhibition of inflammatory reaction or surviving cells stimulation.
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