臺灣博碩士論文加值系統

頭部外傷為全世界及台灣之主要死因之一，與癌症與心腦血管疾病不同，頭部外傷好發於年輕人，致死率極高，即使存活也約有25 % 病患癱瘓、長期失能的情形，耗費社會成本至鉅。然而目前臨床上對於頭部外傷之治療僅為支持性療法，本研究即探討白血球間質素-1（IL-1）在頭部外傷引發神經元死亡所扮演之角色。本論文中頭部外傷之誘發參考Marmarou 等人之動物模式，利用重物（450 g砝碼）自高處落下（2公尺）撞擊大白鼠頭部後，分別在不同時間點（0 hr，3 hr，6 hr，12 hr，48 hr，96 hr，168 hr）將動物麻醉犧牲，利用組織染色法來評估頭部外傷後，大腦皮質（supraventricular gyrus）、海馬迴（CA 3）、紋狀體（cingulate gyrus）等不同部位之神經元傷害程度。結果發現大部份神經元死亡，並且出現顯著的細胞空隙，隨時間增加細胞死亡的現象亦越明顯。以反轉錄酶-聚合酶連鎖反應觀察腦部細胞激素IL-1α，IL-1β之表現，於頭部外傷後的三小時在這些部位皆偵測到IL-1α mRNA（顯著表現達四倍）、IL-1β mRNA（顯著表現達二倍）的表現，而IL-1α mRNA持續表現達一個禮拜；而IL-1β mRNA只持續表現至傷害後的九十六小時。同樣以西方點墨法觀察腦部細胞激素蛋白質之表現，頭部外傷後的三小時即測得IL-1α、β之蛋白質表現（顯著表現達二倍），進一步証實頭部外傷會誘發IL-1α、β之蛋白質的生成。以帶有β-galactosidase的E.coli突變株M4當追蹤物，偵測頭部外傷對於腦-血管屏障開啟之探討，結果顯示，腦血管屏障之開啟在傷害後三小即可看見，且隨時間之延長，腦血管屏障之開啟更加明顯，但在傷後一個禮拜，腦血管屏障癒合。同時由雙重螢光染色結果得知，在頭部外傷後主要表現細胞激素之細胞種類為中樞神經系統中的神經元及星狀細胞。並以免疫組織化學染色法探討血腦屏障對週邊及中樞之影響，結果證實細胞激素會因為血腦屏障的受損而由週邊進到中樞，進一步加劇中樞的發炎反應。並以西方點墨法探討頭部外傷後，細胞激素之訊息傳遞路徑，結果得知頭部外傷後會活化MAPK，且是藉由p-ERK的作用，啟動MAPK訊號傳遞。同時由腦室分別給予anti-IL-1α及anti-IL-1β抗體處理老鼠，依結果發現此兩種抗體均能有效抑制頭部外傷所造成之神經元死亡，進一步証實IL-1在頭部外傷的過程中的確對於神經元的死亡扮演著很重要的影響角色。

Traumatic brain injury (TBI) is one of the most prevalent causes of morbidity and mortality all over the world. It mainly affects young adults in their most productive stage of life and makes an enormous social and economical cost. However, there is no adequate treatment to reduce the cerebral damage following brain injury. This study assessed the effect of IL-1 on TBI-induced neuron damage. TBI model set up by the calibrated weight-drop device (450 g weight, 2 m height) and the rats were divided into sham operation and different time course after TBI (0 hr, 3 hr, 6 hr, 12 hr, 48 hr, 96 hr and 168 hr). The neuronal damage was verified by hematoxylin and eosin staining and microscopic examination. IL-1α and IL-1β mRNA expression were detected by RT-PCR and the protein expression was measured by Western blot. Blood brain barrier permeability during TBI was detected by β-galactosidase and immunofluorescent double labeling was used to elucidate the site of IL-1 release. In this study, we found that IL-1α mRNA expression peaked at 12 hr （increased 8-fold）and lasted for 168 hr, and IL-1β mRNA was significantly expressed after 12 hr（increased 4-fold）after TBI but only lasted for 96 hr. In the protein level, IL-1α and IL-1β level significantly elevated 3 hr（increased 2-fold）after TBI and lasted for 168 hr. The BBB permeability increased 3 hr after TBI and back to normal level about 1 week later. The possible intracellular signal transduction pathway of TBI is mediated by mitogen activated protein kinase phosphorylation. By the immunofluorescent double labeling, we found IL-1α was released from neuron and IL-1β was released from astrocyte. Moreover, administration of IL-1α and IL-1β antibodies significantly protects animals from TBI-induced neuronal loss. Theses results evidenced that IL-1 plays an essential roles in TBI-induced neuron damage and suggested that IL-1α and IL-1β elicited different effect during TBI.

推薦書 ----------------------------------------------------- i
審定書 ----------------------------------------------------- ii
授權書 -----------------------------------------------------iii
中文摘要 --------------------------------------------------- iv
英文摘要 --------------------------------------------------- vi
誌謝 ------------------------------------------------------viii
目錄 ------------------------------------------------------ix
圖目錄 -----------------------------------------------------x
縮寫指引 --------------------------------------------------xiii
緒論 -------------------------------------------------------- 1
實驗設計 --------------------------------------------------- 14
材料及方法 -------------------------------------------------15
結果 ------------------------------------------------------ 29
討論 ----------------------------------------------------- 34
參考文獻 --------------------------------------------------- 43
圖附錄 ---------------------------------------------------- 54

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