Correlação da intensidade do sinal em FLAIR e os níveis de mediadores inflamatórios no hipocampo de pacientes com epilepsia do lobo temporal e esclerose mesial temporal

We investigated a relationship between the FLAIR signal found in mesial temporal sclerosis (MTS) and inflammation. Twenty nine patients were selected through clinical and MRI analysis and submitted to cortico-amygdalo-hippocampectomy to seizure control. Glutamate, TNF&#945;, IL1, nitric oxide (NO) levels and immunostaining against IL1&#946; and CD45 was performed. Control tissues (n=10) were obtained after autopsy of patients without neurological disorders. The glutamate was decreased in the temporal lobe epilepsy (TLE) -MTS group (p<0.001), suggesting increased release of this neurotransmitter. The IL1&#946; and TNF&#945; were increased in the hippocampus (p<0.05) demonstrating an active inflammatory process. A positive linear correlation between FLAIR signal and NO and IL1&#946; levels and a negative linear correlation between FLAIR signal and glutamate concentration was found. Lymphocytes infiltrates were present in hippocampi of TLE patients. These data showed an association between hippocampal signal alteration and increased inflammatory markers in TLE-MTS.Este estudo foi delineado para investigar a presença de relação entre a intensidade de sinal em FLAIR e níveis de citocinas, óxido nítrico (NO) e glutamato no hipocampo de pacientes com epilepsia do lobo temporal refratária, associada com esclerose mesial (TLE-MTS). Vinte e nove pacientes foram selecionados através de análise clínica e de ressonância magnética (RM) que foram submetidos a cortico-amigdalo-hipocampectomia para o controle das crises. Os níveis de glutamato foram avaliados por HPLC, as citocinas TNF&#945; e IL1&#946; por ELISA e os níveis de NO via NO system. Avaliamos também por imuno-histoquímica a expressão de IL1&#946; e CD45 em tecidos controles e com esclerose. Tecido controle foi obtido após autópsia de indivíduos mortos sem disfunções inflamatórias e neurológicas (n=10). A concentração de glutamato se mostrou reduzida no tecido TLE-MTS (p<0,001) sugerindo aumento na liberação desse neurotransmissor. TNF&#945; e IL1&#946; também apresentaram níveis elevados no hipocampo dos pacientes (p<0,05), demonstrando um processo inflamatório crônico. Houve uma correlação linear positiva entre a intensidade do sinal em FLAIR e os níveis de NO e IL1&#946;. Em contraste, uma correlação linear negativa foi encontrada entre a intensidade do sinal em FLAIR e níveis de glutamato no hipocampo com esclerose. Infiltrado linfocitário hipocampal também foi visualizado pela imuno-marcação com CD45 em pacientes com TLE-MTS. Esses dados mostraram uma associação entre alteração de sinal na RM e marcadores inflamatórios em pacientes com TLE-MTS.FAPESP - CInNAPCeCNPqMCT - Instituto Nacional de Neurociência Translaciona

Hippocampal hipersignal on MRI is relacionated with IL1, TNF, Nitric Oxide and glutamate levels

Objetivos: Estudar a existência de correlação linear entre a intensidade do sinal na IRM do hipocampo em FLAIR na ELT com EH e a concentração de GLU, NO, IL1β e TNFα. Em outro enfoque, objetivamos comparar as concentrações hipocampais destes mediadores entre os pacientes com ELT e EH e controles obtidos de autópsia. Metodologia: Estudamos 29 pacientes com ELT por EH e 20 indivíduos controles (10 autópsia e 10 controles de IRM). O sinal na IRM foi medido em FLAIR (Leonardo, Syngo MR 2004A Siemens Medical Solutions) tanto para pacientes como para controles. As concentrações de GLU formam obtidas por cromatografia líquida (HPLC), a de IL1β e TNFα foram obtidas por ELISA (Kit BD Opteia) e a de NO foi aferida pelo equipamento NOATM 280, Sievers Instruments. Os dados foram avaliados utilizando-se os testes t de Student para a comparações de concentrações de mediadores nos diferentes tecidos e o teste exato de Fisher, de Kolmogorov-Smirnov, de Levene para a avaliação da correlação linear de Pearson entre as variáveis. Resultados: Quanto avaliamos as concentrações dos diferentes mediadores em tecido com EH e controle observamos aumento da concentração de IL1β e TNFα e redução do GLU e GABA na EH, sem alterações significativas do NO. Houve correlação linear positiva entre as concentrações de NO, e hipersinal em FLAIR tanto na cabeça, corpo e cauda do hipocampo. A citocina IL1β mostrou correlação linear positiva com o sinal em FLAIR na cauda hipocampal. Além disso, houve correlação linear inversa entre o sinal de FLAIR e a concentração de GLU no hipocampo de pacientes com EH. Conclusão: Esses dados sugerem haver correlação entre liberação de GLU e intensidade de sinal em FLAIR na IRM. Por outro lado a correlação positiva entre NO e IL1β e sinal em FLAIR também sugere associação com o fenômeno inflamatório e quantidade de água no hipocampo. Desta forma, concluímos que na EH há inflamação e excitotoxicidade, mesmo nas fases crônicas da síndrome, flagradas pelas alterações das concentrações de IL1β, TNFα e GLU e NO. Estas alterações podem estar correcionadas com o hipersinal da IRM em FLAIRPurpose: To study the existence of linear correlation between the intensity of the MRI T2 signal in the hippocampus of patients with temporal lobe epilepsy (TLE) and hippocampal sclerosis (HS) and the concentration of GLU, NO, TNFα and IL1β. In another approach, we aimed to compare the hippocampal concentrations of these mediators between patients with TLE with HE and control subjects, obtained from autopsy. Methods: We studied 29 patients with TLE with HS and 20 control subjects (10 autopsy and 10 controls for MRI). The signal was measured on MRI in T2 (Leonardo, 2004a MR Syngo Siemens Medical Solutions) for both patients and for controls. The concentrations of GLU are obtained by liquid chromatography (HPLC), the IL1β and TNFα concentration were obtained by ELISA (BD Opt Kit) and the NO level was measured by the equipment NOATM 280, Sievers Instruments. Data were evaluated using the Student t test for the comparison of mediator’s concentrations in the hippocampus of control and patients with HS and the Fisher exact test, the Kolmogorov-Smirnov, Levene was employed for the evaluation of the Pearson's linear correlation between variables.Results: The evaluation of the inflammatory mediator’s concentration in tissues with HS and controls showed increased concentration of IL1β and TNFα as well as reduction of GLU and GABA in HS, without significant changes of NO levels. A positive linear correlation between NO levels and signal on T2 in the head, body and tail of hippocampus were also found. The cytokine IL1β levels showed positive 160linear correlation with the MRI T2 signal with the tail of the hippocampus. Moreover, an inverse linear correlation between the T2 MRI signal and the concentration of GLU in the hippocampus of patients with HE was also detected. Conclusion: These data suggest a correlation between the increased release of GLU and the intensity of T2 MRI signal. Furthermore the positive correlation between NO and IL1β and T2 MRI signal suggests an association between hipersignal, inflammation and amount of water in the hippocampus of these patients. Thus, we conclude that there is inflammation and excitotoxicity in TLE with HS, even in chronic stages of this syndrome, visualized by changes in the concentrations of IL1β, TNFα, GLU and NO. These changes could be related to the hypersignal on T2 MRI.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundo de Auxílio aos Docentes e Alunos (FADA)Instituto Nacional de Neurociência Translacional (INNT)TEDEBV UNIFESP: Teses e dissertaçõe

Kinin B1 and B2 receptors are overexpressed in the hippocampus of humans with temporal lobe epilepsy

Molecular biology tools have been employed to investigate the participation of peptides in human temporal lobe epilepsy ME). Active polypeptides and their receptors have been related to several brain processes, such as inflammation, apoptosis, brain development, K+ and Ca2+ channels' activation, cellular growth, and induction of neuronal differentiation. Previous works have shown a neuroprotector effect for kinin B2 receptor and a deleterious, pro-epileptogenic action for kinin B1 receptor in animal models of TLE. the present work was delineated to analyze the kinin B1 and B2 receptors expression in the hippocampus of patients presenting refractory mesial TLE. the hippocampi were removed during the patients surgery in a procedure used for seizure control and compared with tissues obtained after autopsy. Nissl staining was performed to study the tissue morphology and immunohistochemistry, and Western blot was used to compare the distribution and levels of both receptors in the hippocampus. in addition, real time PCR was employed to analyze the gene expression of these receptors. Nissl staining showed sclerotic hippocampi with hilar, granular, and pyramidal cell loss in TLE patients. Immunohistochemistry and Western blot analyses showed increased expression of kinin B1 and B2 receptors but the real-time PCR data demonstrated increased mRNA level only for kinin B2 receptors, when compared with controls. These data show for the first time a relationship between human TLE and the kallikrein-kinin system, confirming ours previous results, obtained from experimental models of epilepsy. (c) 2006 Wiley-Liss, Inc.Universidade Federal de São Paulo, Dept Expt Neurol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Patol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biofis, São Paulo, BrazilFCMUSP, Dept Anat Patol Incor, São Paulo, BrazilUniversidade Federal de São Paulo, Disciplina Neurol Clin, Dept Neurol Neurocirurgia, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Diagnost Imagem, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Bioquim, São Paulo, BrazilCtr Univ Nove de Julho, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Expt Neurol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Patol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biofis, São Paulo, BrazilUniversidade Federal de São Paulo, Disciplina Neurol Clin, Dept Neurol Neurocirurgia, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Diagnost Imagem, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Bioquim, São Paulo, BrazilWeb of Scienc

The renin-angiotensin system is upregulated in the cortex and hippocampus of patients with temporal lobe epilepsy related to mesial temporal sclerosis

Purpose: As reported by several authors, angiotensin II (AngII) is a proinflammatory molecule that stimulates the release of inflammatory cytokines and activates nuclear factor kappa B (NF kappa B), being also associated with the increase of cellular oxidative stress. Its production depends on the activity of the angiotensin converting enzyme (ACE) that hydrolyzes the inactive precursor angiotensin I (AngI) into AngII. It has been suggested that AngII underlies the physiopathological mechanisms of several brain disorders such as stroke, bipolar disorder, schizophrenia, and disease. the aim of the present work was to localize and quantify AngII AT1 and AT2 receptors in the cortex and hippocampus of patients with temporal lobe epilepsy related to mesial temporal sclerosis (MTS) submitted to corticoamygdalohippocampectomy for seizure control.Method: Immunohistochemistry, Western blot, and real-time PCR techniques were employed to analyze the expression of these receptors.Results: the results showed an upregulation of AngII AT1 receptor as well as its messenger ribonucleic acid (mRNA) expression in the cortex and hippocampus of patients with MTS. in addition, an increased immunoexpression of AngII AT2 receptors was found only in the hippocampus of these patients with no changes in its mRNA levels.Discussion: These data show, for the first time, changes in components of renin-angiotensin system (RAS) that could be implicated in the physiopathology of MTS.Universidade Federal de São Paulo, Dept Neurol & Neurosurgery, Discipline Expt Neurol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Neurol & Neurosurgery, Discipline Nephrol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Pathol, São Paulo, BrazilUniv São Paulo, Sch Med, Dept Pathol, São Paulo, BrazilUniversidade Federal de São Paulo, Discipline Clin Neurol, Dept Biochem, São Paulo, BrazilUniversidade Federal de São Paulo, Discipline Clin Neurol, Dept Neurol & Neurosurg, São Paulo, BrazilNove Julho Univ, Dept Sci Rehabil, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Neurol & Neurosurgery, Discipline Expt Neurol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Neurol & Neurosurgery, Discipline Nephrol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Pathol, São Paulo, BrazilUniversidade Federal de São Paulo, Discipline Clin Neurol, Dept Biochem, São Paulo, BrazilUniversidade Federal de São Paulo, Discipline Clin Neurol, Dept Neurol & Neurosurg, São Paulo, BrazilWeb of Scienc