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研究生:劉為麟
研究生(外文):Wei-Lin Liu
論文名稱:糖皮質固醇類藥物調控活化態星形膠質細胞分泌蛋白多醣的能力及影響神經纖維生長之探討
論文名稱(外文):Regulation of methylprednisolone on the expression of chondroitin sulfate proteoglycans in reactive astrocytes and the neurite outgrowth in dorsal root ganglion neurons
指導教授:楊維中楊維中引用關係
學位類別:碩士
校院名稱:臺北醫學大學
系所名稱:生物醫學材料研究所
學門:工程學門
學類:生醫工程學類
論文種類:學術論文
論文出版年:2005
畢業學年度:93
語文別:中文
中文關鍵詞:糖皮質固醇蛋白多醣細胞外間質星形膠質細胞
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中樞神經系統在受傷後軸突再生的能力有限,往往會造成病患神經功能的永久喪失,其中存在於細胞外間質(extracellular matrix, ECM)之硫酸軟骨素蛋白多醣(chondroitin sulfate proteoglycans, CSPGs)在受傷後會由再活化的星形膠質細胞所分泌,且存在於受傷部位周圍形成神經膠疤痕組織被認為是抑制軸突再生的因子之一。目前常用於治療中樞神經傷害的糖皮質固醇藥物具有神經保護的作用,但對於此類藥物是否會影響星形膠質細胞之活化及分泌細胞外間質的能力並不是很清楚。本研究即利用體外培養初代星形膠質細胞並給予興奮性氨基酸麩胺酸相似 物R,Sα-amino-3-hydroxyl-5-methylisoxazole-4- propionic acid (AMPA)的刺激以模擬在體內星狀膠質細胞受傷的情況,並觀察星形膠質細胞在此情況下分泌細胞外間質成分的能力是否會受到糖皮質固醇藥物,methylprednisolone (MP)所影響,並同時探討受傷後的神經膠質細胞所分泌的細胞外間質成分對於神經軸突生長的影響。
本研究結果顯示MP的加入的確可以有效抑制硫酸軟骨素(chondroitin sulfate, CS)氨基葡聚醣(glycosaminoglycan, GAG)以及 hylauronan-binding類CSPGs在受傷模式下的表現,但對於small leucine-rich這類的CSPGs則效果較不明顯。近一步的分子基因調控研究顯示MP對於hylauronan-binding類CSPGs之抑制作用可能是透過糖皮質固醇接受器(glucocorticoid receptors, GRs)的作用,且給予MP的星形膠質細胞培養基可以促進神經軸突的生長或許是透過對於ECM中CSPGs的抑制作用。另外觀察不同類之CSPGs對於背根神經節神經細胞軸突的生長的影響,研究結果顯示CSPG中GAG及core protein對於神經軸突的生長有不同程度的影響且其調控可能是透過不同的作用機轉,這些結果可以推測MP類藥物參與了調控中樞神經受傷後細胞外間質的重組並藉以影響受傷神經軸突的生長。
中文摘要…………………………………………………………..1
英文摘要…………………………………………………………..3
第一章 文獻回顧………………………………………………..5
一、星形膠質細胞(astrocyte)……………………………...6
二、細胞外間質(extracellular matrix, ECM)………………8
三、蛋白多醣(proteoglycan)……………………………….9
四、背根神經節神經細胞(dorsal root ganglion neuron)…..11
五、糖皮質固醇類藥物(glucocorticoid)…………………...12
第二章 實驗目的………………………………………………...15
第三章 實驗設計………………………………………………...16
實驗一:初代星形膠細胞細胞培養………………………………16
實驗二:星狀膠質細胞活性分析…………….…………………...18
實驗三:圓點點墨法……………………………………...……….19
實驗四:蛋白質萃取……………….…………………...…………21
實驗五:西方點墨法……………...…………………...….…….....23
實驗六:反轉錄聚合酶鍊鎖反應…………………..……..………25
實驗七:轉錄因子活性實驗………………………………………26
實驗八:初代背根神經節神經細胞培養……………………........27
實驗九:細胞螢光免疫染色..………………………………..........31
實驗十:統計分析方法…...………………………………….........32
第四章 實驗結果…………………………………….…….........33
第五章 討論……….………………………………...…………..38
第六章 結論……………………………………………………..43
第七章 未來研究方向…………………………………………..44
第八章 參考文獻………………………………………………..45
第九章 實驗圖表………………………………………………..53
第十章 附錄……………………………………………………..77
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