臺灣博碩士論文加值系統

過去研究顯示，臍帶血含有低於0.2％的造血幹細胞（CD34+），在液體撞擊腦創傷（FPI）的動物實驗模式，具有減少神經受損的功效。本研究主要是純化臍帶血的CD34+細胞（大於95%），研究幹細胞改善FPI大鼠的機轉。實驗大鼠共分三組：（1）對照組；（2）FPI並移植CD34-細胞 （5x105臍帶血淋巴球與單核球，含有0.2％CD34+細胞）；（3）FPI並移植CD34+細胞（5x105臍帶血淋巴球與單核球，含有95％CD34+細胞）。FPI四天後評估實驗大鼠的行為障礙與腦梗塞、發炎、細胞凋亡、血管生成和神經再生。與對照組相比，CD34-細胞治療FPI大鼠，仍有運動與認知障礙、腦梗塞、細胞凋亡和發炎現象。藉由CD34+細胞治療，可以明顯減緩FPI導致的神經損傷與腦梗塞、細胞凋亡和發炎現象。此外，CD34+細胞可以移至受損腦部，有效地促使受損區域的血管新生與神經再生。
本研究結果顯示，純化臍帶血CD34+造血幹細胞，對於減緩創傷性腦損傷的大鼠，可能具有療效。

Umbilical cord blood contained <0.2% CD34+ cells have been shown to be beneficial in reducing neurological deficits in animals after fluid percussion injury（FPI）. This study aimed to generate cord blood-derived CD34+ cells（>95%）and to investigate the mechanisms underlying their beneficial effects in treating FPI in rats. Rats were divided into three groups:（1）sham operation; （2）FPI+CD34- cells（5×105 cord blood lymphocytes and monocytes that containing <0.2% CD34+ cells）; and （3） FPI+CD34+ cells（5×105 cord blood lymphocytes and monocytes that contained 95% CD34+ cells）. Behavioral dysfunction and brain infarction, inflammation, apoptosis, angiogenesis, and neurogenesis were evaluated 4 days post FPI. As compared to sham operation controls, CD34- -treated FPI rats had motor and cognitive dysfunctions and cerebral infarction, apoptosis, and inflammation. FPI-induced neurological dysfunction and cerebral infarction, apoptosis, and inflammation could be significantly attenuated by CD34+ cell therapy. In addition, CD34+ cells migrated to the injured brain regions and significantly promoted both angiogenesis and neurogenesis in the injured brain.
The results indicate that therapy using umbilical cord blood-derived CD34+ cells may be beneficial in attenuating traumatic brain injury in rats.

中文摘要…………………………………………………………………I
英文摘要 ……………………………………………………………… II
誌謝 …………………………………………………………………… IV
目錄 …………………………………………………………………… V
圖目錄………………………………………………………………… VIII
表目錄………………………………………………………………… XI
常用縮寫及專有名詞………………………………………………… XII
第一章 緒論 …………………………………………………… 1
第一節 創傷性腦損傷………………………………………… 4
第二節 幹細胞介紹…………………………………………… 6
第三節 大腦海馬介紹………………………………………… 12
第四節 研究目的……………………………………………… 14
第二章 材料與方法…………………………………………… 15
第一節 實驗材料……………………………………………… 15
第一項 實驗藥品……………………………………………… 15
第二項 實驗儀器……………………………………………… 16
第三項 實驗動物……………………………………………… 19
第二節 實驗方法……………………………………………… 20
第一項 手術…………………………………………………… 20
第二項 生理參數監測………………………………………… 22
第三項 微透析測量大腦海馬回之麩氨酸、甘油、乳酸鹽/焦葡萄酸鹽和一氧化氮代謝
物………………… 23
第四項 人類造血幹細胞之準備……………………………… 24
第五項 實驗分組……………………………………………… 25
第六項 實驗過程……………………………………………… 26
第七項 運動功能的評估……………………………………… 28
第八項 認知功能的評估……………………………………… 29
第九項 內皮前驅細胞………………………………………… 30
第十項 腦梗塞評估…………………………………………… 31
第十一項 凋亡細胞之凋亡檢測………………………………… 32
第十二項 溴脫氧尿嘧啶核苷標記……………………………… 32
第十三項 免疫組織化學染色…………………………………… 33
第十四項 細胞激素的測定……………………………………… 35
第十五項 統計分析……………………………………………… 36
第三章 實驗結果……………………………………………… 37
第一節 CD34+細胞改善FPI大鼠所引發的顱內壓升高和腦部血
灌注降低……………………………………………… 37
第二節 CD34+細胞改善FPI大鼠所引發的腦缺血和神經元損
傷……………………………………………………… 39
第三節 CD34+細胞改善FPI引發的運動和認知功能障礙…… 41
第四節 CD34+細胞改善FPI引發的腦梗塞…………………… 45
第五節 CD34+細胞改善FPI引發的腦神經細胞凋亡………… 47
第六節 CD34+細胞治療FPI刺激血管生成…………………… 52
第七節 CD34+細胞治療FPI刺激神經再生…………………… 56
第八節 CD34+細胞改善FPI引發的炎症反應………………… 61
第九節 CD34+細胞治療FPI提高循環內皮先祖細胞………… 63
第十節 經由免疫組織化學染色測定CD34+細胞到達FPI損傷
區域…………………………………………………… 65
第四章 討論…………………………………………………… 67
第五章 結論…………………………………………………… 72
第六章 參考文獻……………………………………………… 73
英文部分……………………………………………… 73
中文部分……………………………………………… 86

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