Perosa Sandra Regina, Argañaraz Gustavo Adolfo, Goto Eduardo Massatoshi, Costa Luciana Gilbert Pessoa, Konno Ana Carla, Varella Pedro Paulo Vasconcellos, Santiago Joselita Ferreira Carvalho, Pesquero João Bosco, Canzian Mauro, Amado Debora, Yacubian Elza Marcia, Carrete Henrique, Centeno Ricardo Silva, Cavalheiro Esper Abrão, Silva Jose Antonio, Mazzacoratti Maria da Graça Naffah
Departamento de Neurologia Experimental, Universidade Federal de São Paulo, Sao Paulo, Brazil.
Hippocampus. 2007;17(1):26-33. doi: 10.1002/hipo.20239.
Molecular biology tools have been employed to investigate the participation of peptides in human temporal lobe epilepsy (TLE). Active polypeptides and their receptors have been related to several brain processes, such as inflammation, apoptosis, brain development, K(+) and Ca(2+) 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.
分子生物学工具已被用于研究肽在人类颞叶癫痫（TLE）中的作用。活性多肽及其受体与多种脑过程有关，如炎症、细胞凋亡、脑发育、钾离子（K⁺）和钙离子（Ca²⁺）通道的激活、细胞生长以及神经元分化的诱导。先前的研究表明，在TLE动物模型中，激肽B2受体具有神经保护作用，而激肽B1受体具有有害的促癫痫作用。本研究旨在分析难治性内侧TLE患者海马中激肽B1和B2受体的表达。在患者手术过程中切除海马，该手术用于控制癫痫发作，并与尸检后获得的组织进行比较。采用尼氏染色研究组织形态学，免疫组织化学和蛋白质印迹法用于比较海马中两种受体的分布和水平。此外，实时定量聚合酶链反应（real time PCR）用于分析这些受体的基因表达。尼氏染色显示TLE患者海马硬化，伴有门区、颗粒细胞和锥体细胞丢失。免疫组织化学和蛋白质印迹分析显示激肽B1和B2受体表达增加，但与对照组相比，实时定量聚合酶链反应数据仅显示激肽B2受体的mRNA水平增加。这些数据首次表明人类TLE与激肽释放酶-激肽系统之间的关系，证实了我们先前从癫痫实验模型中获得的结果。
