Serum levels of matrix metalloproteinase 2 and its inhibitor after tonic-clonic seizures.

Damage to the blood-brain barrier (BBB) may result from on-going neuroinflammation, which can lead to leakage of blood components, such as leukocytes and serum proteins, into the brain, resulting in disturbed brain homeostasis. The aim of the project was to examine the involvement of modulatory proteins in the processes of BBB integration after epileptic seizures. We investigated serum changes in the levels of MMP-2 and MMP-7 and its inhibitors after seizures in epilepsy patients. Concentrations of these proteins were measured by ELISA in 50 patients at 1-3, 24, and 72 h after generalized tonic-clonic seizures and once in participants of the control group. The level of MMP-2 in serum was slightly higher after seizures (at 1-3 h time point), but the difference was not statistically significant. The levels of trombospondine (TSP) - 1 and - 2 were decreased at 1-3 h after seizures. The expression of TIMP-2 was increased 1 and 24 h after seizures. There were no significant changes in the level of α2-macroglobulin and MMP-7. Changes in the expression of both specific and non-specific inhibitors indicate the initiation of repair processes of the blood-brain barrier and improvement of its integrity. Since we performed serum analysis, further studies are necessary to investigate the correlation with the expression of the investigated markers in the brain. Perhaps this will allow for the identification of new biomarkers associated with epileptic seizures.

Neurodegeneration and inflammation in hippocampus in experimental autoimmune encephalomyelitis induced in rats by one--time administration of encephalitogenic T cells.

Cognitive dysfunction is relatively frequent in multiple sclerosis (MS) and it happens from the early stages of the disease. There is increasing evidence that the grey matter may be involved in autoimmune inflammation during relapses of MS. The purpose of this study was to evaluate if a single transfer of encephalitogenic T cells, mimicking a relapse of MS, may cause hippocampal damage and memory disturbances in rats. Lewis rats were injected with anti-MBP CD4+ T cells, that induced one-phase autoimmune encephalomyelitis (EAE) with full recovery from motor impairments at 10-15 days. The spatial learning and memory were tested by the Morris water maze test in control and EAE animals, 30 and 90 days post-induction (dpi). The neural injury and inflammation was investigated in the hippocampus by immunohistochemistry and quantitative analyses. There was a marked decrease in the number of CA1 and CA4 pyramidal neurons 5 dpi. The loss of neurons then aggravated till the 90 dpi. An increase in microglial and astroglial activation and in pro-inflammatory cytokines mRNA expression in the hippocampus, were present 30 and 90 dpi. Nerve growth factor and brain-derived neurotrophic factor mRNA levels were also significantly elevated. The water maze test, however, did not reveal memory deficits. The present data indicate that a single transfer of autoimmune T cells results in preserved inflammation and probable on-going neuronal injury in the hippocampus, long after recovery from motor disturbances. These findings suggest that any relapse of the MS may start the neurodegenerative process in the hippocampus, which is not necessarily connected with memory deficits.

The impact of age and gender on the striatal astrocytes activation in murine model of Parkinson's disease.

OBJECTIVE: The aim of the present study was to determine how aging and gender influence the response of astrocytes to 1-methyl-4-phenyl-1,2,3,6-tetrahydropiridine (MPTP) intoxication. MATERIALS AND METHODS: To asses the MPTP-induced astrocytes activation in nigro-striatal system, we measured the temporal changes in mRNA and protein expression of the specific astrocytic marker, glial fibrillary acidic protein (GFAP; by RT-PCR and Western blot), in the striatum of male and female C57BL/6 mice (2 and 12-month old) after 6 h and 1, 3, 7, 14 and 21 days post-intoxication. RESULTS: We observed the increases of GFAP mRNA level post-MPTP intoxication in both young and aging males only at early time points, whereas in females (both ages) also at later time points. We noticed maximal increase of GFAP protein content on the 3rd day post-intoxication in young and aged males, whereas in females at the 7-daytime point. CONCLUSIONS: The present results provide additional information of potential relevance to understand the mechanisms of gender and age-related difference in susceptibility of nigro-striatal system to MPTP insult.

Decreased inflammation and augmented expression of trophic factors correlate with MOG-induced neuroprotection of the injured nigrostriatal system in the murine MPTP model of Parkinson's disease.

The response of the immune system during injury of the central nervous system may play a role in protecting neurons. We have previously reported that immunization with MOG 35-55 prior to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced injury of the dopaminergic system promotes less dopamine depletion and less dopaminergic damage of neurons in mice. In this study, we evaluate the influence of MOG immunization on the inflammatory reaction that occurs at the place of injury. C57Bl male mice, 2 and 12 months old, received i.p. injections of MPTP (40 mg/kg) and some groups animals also received an additional injection with myelin oligodendrocyte glycoprotein (MOG) 35-55 in CFA 6 days before MPTP administration. MPTP caused a common inflammatory reaction characterized by microglial activation, infiltration of T cells into the substantia nigra and striatum and increased expression of mRNA encoding pro-inflammatory cytokines (IL-1 beta, TNFalpha, INF gamma) and trophic factors (TGFbeta, GDNF). MOG immunization prior to MPTP administration significantly diminished the microglial reaction and reduced the levels of infiltrating CD8+ lymphocytes. The number of CD4+ T cells remained at the same level as in the MPTP group. Expression of pro-inflammatory cytokines was diminished. The mRNA expression of GDNF was significantly higher in the MOG pretreated mice relative to the MPTP group, both in the 2 month old and 12 month old groups. Since MOG immunization prior to MPTP intoxication appears to prevent nigrostriatal injury, the observed decrease of inflammation and increase of GDNF mRNA expression in the injured areas might represent one of the mechanisms of observed neuroprotection.