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The purpose of this study was to evaluate the action of two types of stressors in Sprague-Dawley (S-D) and Lewis (LEW) rats differing in their hypothalamic-pituitary-adrenal axis activity on locomotion and rearing in an open space. Exposure to restraint immobilization alone (IMO) or this immobilization combined with cold water (22 °C) immersion (IMO+C) lasted for 1 h and started 2 or 5 h before the test. To evaluate the acute and persisting effects of both stressors, four trials were performed in one-week intervals; rats were exposed to the stressors in trial 1 and 3. While in LEW rats both acute stressors produced reduction of locomotion and rearing in all stressed groups, S-D rats responded with a decrease of both parameters only after IMO+C presented 2 h before testing. Neither strain displayed a changed performance one week after the first stress exposure. One week after the second stress exposure rats of both strains exhibited a tendency to an increase of both parameters reaching the significance in several experimental groups. The findings indicate: a) the IMO+C produced stronger behavioral alteration than IMO alone; b) the behavioral responses to stressors were more pronounced in LEW compared to S-D strain; c) change from the reduction of activity to its increase may be interpreted as bi-directional manifestation of long-term effects of immobilization stress.
Experimental autoimmune encephalomyelitis (EAE) is an animal model that mimics many aspects of multiple sclerosis (MS). Chronic or relapsing inflammation of the central nervous system results in the destruction of myelin sheath and cytokines play an important role in the pathogenesis of both MS and EAE. Myelin, oligodendrocytes and neurons are lost due to an inflammatory attack by leukocytes infiltrating the central nervous system (CNS) and releasing cytotoxic cytokines, anti CNS antibodies and large amounts of the excitatory neurotransmitter glutamate. Pharmacological studies have suggested that glutamate receptors mediate white matter injury in a variety of CNS diseases, including multiple sclerosis (MS). Memantine and amantadine are ionotropic glutamate receptors (iGluRs) antagonists. Memantine, a clinically applied drug with N-methyl-D-aspartate (NMDA) receptor antagonistic effects, dose-dependently ameliorates neurological deficits in Lewis rats subjected to experimental autoimmune encephalomyelitis (EAE). The aim of the present study was to investigate the effects of memantine and amantadine on the expression of proinflammatory cytokines such interleukin 1beta (IL-1β), interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α) and various chemokines in the brain of EAE rats. Real-time Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Western Blot were used to analyze the cytokine profile. We noticed increased expression of array of cytokines in experimental group when compared to the control. Dramatic increase of IL-1β, IL-6, TNF-α, and chemokines concentration corresponding to the intensity of neurological symptoms and loss of weight was observed in EAE rats. Administration of iGluR antagonists at an advanced stage of unremitting EAE resulted in amelioration of the disease. Cytokine analysis revealed that memantine significantly decreased the expression of interleukins: IL-6 (65%), IL-1β (60%) and TNF-α (45%) whereas treatment with amantadine reduced only the expression of IL-6 (60%) and TNF-α (15%) when compared to EAE animals. These results show that antagonists of iGlu receptors modulate the course of the disease by reducing the expression of proinflammatory cytokines thereby confirming the involvement of glutamate receptors into pathological mechanisms operating during EAE. This study was supported by grant nr NN401620038 from Polish Ministry of Science and Higher Education
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