do Carmo Cunha Jinger, de Freitas Azevedo Levy Beatriz, de Luca Bianca Aparecida, de Andrade Michele Schultz Ramos, Gomide Vânia Canterucci, Chadi Gerson
Neuroregeneration Center, Department of Neurology, University of São Paulo School of Medicine, University of São Paulo, São Paulo, Brazil.
Wound Repair Regen. 2007 Jan-Feb;15(1):134-46. doi: 10.1111/j.1524-475X.2006.00194.x.
This paper demonstrates glial reaction and changes in the S100beta protein and basic fibroblast growth factor (bFGF, FGF-2) in the border and in the adjacent preserved tissue of the rat spinal cord after a contusion. In view of the expression of FGF-2 and S100beta in reactive glial cells and their ability to promote gliogenesis and neuronal trophism, the molecules have been considered to participate in the wound repair and regenerative events after nervous tissue injury. Adult rats were submitted to a moderate spinal cord (10th thoracic level) contusion induced by a New York University Impactor by dropping a 10 g rod from a distance of 25 mm onto the dorsal surface of the exposed dura spinal cord. Impactor curves and parameters were used to monitor the severity of the trauma. Control rats were submitted to sham operation. The motor behavioral spontaneous recovery was demonstrated by means of a BBB test and the combining behavior score up to 3 weeks after injury. Animals were killed 72 hours, 2, and 3 weeks after surgery and spinal cords were processed for immunohistochemistry to show glial fibrillary acidic protein positive astrocytes and OX-42-positive microglia/macrophages as well as changes in the S100beta and FGF-2 in the border and in the adjacent preserved tissue of the lesioned cords. The changes in the immunoreaction products were quantified by means of morphometric/microdensitometric image analysis, and the cell type expressing S100beta and FGF-2 was analyzed by means of two-color immunofluorescence procedures. Massive increases of S100beta and FGF-2 were found in reactive astrocytes, not in reactive microglia, in the border and in the white and gray matters of adjacent preserved tissue of the contused spinal cord in the periods studied. The results are discussed in view of possible paracrine trophic actions of the reactive astrocytes, mediated by S100beta and FGF-2, triggering wound repair events in the border of the trauma, and also leading to neurotrophism and neuronal plasticity in the adjacent regions. These cellular and molecular responses may interfere with the pattern of behavioral recovery after a contusion injury of the spinal cord.
本文展示了大鼠脊髓挫伤后,损伤边界及相邻保留组织中的胶质反应以及S100β蛋白和碱性成纤维细胞生长因子(bFGF,FGF-2)的变化。鉴于FGF-2和S100β在反应性胶质细胞中的表达及其促进胶质细胞生成和神经元营养的能力,这些分子被认为参与了神经组织损伤后的伤口修复和再生过程。成年大鼠通过纽约大学撞击器造成中度脊髓(胸10水平)挫伤,将一根10克的杆从25毫米的高度落到暴露的硬脊膜脊髓背表面。利用撞击器曲线和参数监测创伤的严重程度。对照大鼠进行假手术。通过BBB测试和联合行为评分来证明损伤后长达3周的运动行为自发恢复情况。在术后72小时、2周和3周处死动物,对脊髓进行免疫组织化学处理,以显示胶质纤维酸性蛋白阳性星形胶质细胞和OX-42阳性小胶质细胞/巨噬细胞,以及损伤脊髓边界和相邻保留组织中S100β和FGF-2的变化。通过形态计量/显微密度图像分析对免疫反应产物的变化进行定量,并通过双色免疫荧光程序分析表达S100β和FGF-2的细胞类型。在所研究的时间段内,在挫伤脊髓相邻保留组织的边界、白质和灰质中,反应性星形胶质细胞而非反应性小胶质细胞中发现S100β和FGF-2大量增加。结合反应性星形胶质细胞可能通过S100β和FGF-2介导的旁分泌营养作用,引发创伤边界的伤口修复事件,并导致相邻区域的神经营养和神经元可塑性,对这些结果进行了讨论。这些细胞和分子反应可能会干扰脊髓挫伤损伤后的行为恢复模式。