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    題名: 探討在大鼠脊髓半切損傷動物模式中蛋白多醣的表現
    Expressions of proteoglycans in rat with spinal cord hemisection injury
    作者: 張永恆
    Chang, Yung-Heng
    貢獻者: 楊維中
    關鍵詞: 脊髓半切損傷;蛋白多醣;透明質酸酶;-4;硫酸軟骨素蛋白多醣;C6 神經膠細胞
    spinal cord injury;proteoglycan;hyaluronidase - 4;chondroitin sulfate proteoglycan;C6 glioma
    日期: 2012-07-12
    上傳時間: 2019-06-12 12:20:37 (UTC+8)
    摘要: 中樞神經系統在受傷後其神經軸突自我再生及修復的能力有限，往往會造成傷者神經永久的損傷導致許多功能的喪失。根據過去的研究顯示，導致軸突無法再生修復的主要原因為受傷部位所形成的神經膠質疤痕，此疤痕組織中存在著許多細胞外間質 (Extracellular matrix, ECM)，主要的成分為蛋白多醣 (Proteoglycans) 其中的硫酸軟骨素蛋白多醣 (Chondroitin sulfate proteoglycans, CSPGs) 為目前所知最主要抑制神經軸突再生的物質，然而這些蛋白多醣對於神經軸突並不完全是抑制生長的物質，也有些具有促進神經軸突再生的作用，而近年來發現體內會分泌一種內生性的水解酶 －透明質酸酶 (Hyaluronidase)，可將硫酸軟骨素蛋白多醣分解，但在中樞神經系統受傷後之表現並不清楚。本研究的目的在分析中樞神經系統受傷後，存在於血清及腦脊髓液中蛋白多醣及其分解酵素之表現量，評估其是否可以當作反映神經再生及功能性恢復情況的指標。
    　　本實驗使用六週齡 Sprague-Dawley (SD) 大鼠，隨機分成二組，分別為控制組 (僅將脊椎T9部位的椎間板去除)、SCI (Spinal cord injury) 組 (將脊椎T9部位的椎間板去除後給予橫切)。手術後每週利用BBB運動評分量表 (Basso, Beattie and Breanahan locomotor scale) 評估大鼠運動功能的恢復，並取其血清及腦脊髓液 (Cerebrospinal fluid, CSF) 以點漬法 (Dot blot) 分析蛋白多醣及酵素Hyaluronidase-4之含量濃度，並於手術後六週進行腳印拓印 (Footprint) 及膀胱功能評估並犧牲，取其脊髓檢體進行組織切片分析。
    　　本研究分析受傷後蛋白多醣及酵素表現量的變化與其受傷狀況，及運動與膀胱功能恢復狀況之關聯性。有助於了解特定蛋白多醣及其分解酵素是否可作為脊髓半切受傷情況評估之生物標記及其臨床應用。
    After spinal cord injury (SCI), the reactive glial cells secrete increased amount of proteoglycans (PGs) during the wound healing process. Overproduced proteoglycans may form astroglial scars, which blocks axonal regeneration and causes the impaired neuronal function repair. Chondroitin sulfate proteoglycans (CSPGs), a type of the protepglycans, are known to inhibit axonal regeneration. In contrast, heparin sulfate proteoglycans (HSPGs) may promote neurite outgrowth. In recent years, a hydrolysis enzyme, hyaluronidases-4, has been considered to be the an endogenous enzyme acting at the initial step in the catabolism of chondroitin sulfate. However, the expression and distribution of hyaluronidase-4 (Hyal4) after CNS injury is unclear. In this study, we followed the expressions of various types of proteoglycans and their catabolic enzymese in a rat model with spinal cord hemisection injury for 6 weeks. Six-week-old Sprague-Dawley (SD) rats were randomly divided into 2 groups including control (laminectomy at T9) and spinal cord injury (laminectomy and hemisection at T9). Serum and cerebrospinal fluid samples were collected every week. The expressions of the proteoglycans and the CSPG catabolic enzyme, Hyal-4 were analyzed by dot blot analyses. Immunohistochemistry was carried out for the proteoglycans and the enzumes in spinal cord tissue. The altered expressions of the proteoglycans and Hyal-4 were correlated with the severity of SCI based on the BBB score and the motor function..
    The altered expressions of PGs and hyaluronidae-4 maybe used as biomarkers to evaluate the severety and the functional recovery after SCI.
    描述: 碩士
    指導教授-楊維中
    委員-李怡萱
    委員-蔡翠敏
    資料類型: thesis
    顯示於類別:[生醫材料暨組織工程研究所] 博碩士論文

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