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研究生:蔡秀貞
研究生(外文):Hsiu-chen Tsai
論文名稱:血小板製備過程對其活性與功能之影響
論文名稱(外文):A study of the effects of preparation on the activation and function of platelets
指導教授:陳和瑟陳和瑟引用關係
指導教授(外文):Angela Chen
學位類別:碩士
校院名稱:國立中山大學
系所名稱:生物醫學科學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:153
中文關鍵詞:血小板凝集流體細胞儀單一捐贈者血小板
外文關鍵詞:single-donor plateletsflow cytometryplatelet aggregation
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血小板引起血管的收縮及血塊的形成首先是靠著血小板的活化,活化後的訊息傳遞會使血小板的表面滿佈蛋白質分子,如血小板表面醣蛋白P-選擇素 (P-selectin, CD62P) 等。這些表面醣蛋白在活化的血小板表面會形成醣蛋白複合體,如果再結合上細胞外受體,如膠原及凝血酶等後可以產生凝集。活化的血小板亦可以使促凝劑,如花生四烯酸、二磷酸腺苷、腎上腺素、血清素、纖維蛋白原、von Willebrand factor (vWF) 等釋放,更進一步活化鄰近的血小板。血液中心之標準的血小板分離流程是先將自分離機上取下的血小板靜置於室溫中一小時後,再放置於固定速率的震盪器上搖盪儲存;但有些醫院則省略靜置一小時的動作,直接將自分離機上取下的血小板放置於固定速率的震盪器上搖盪儲存。目前並不清楚對於省略掉血小板搖盪儲存前的靜置一小時對血小板活化情形及功能是否有影響。本計畫以流體細胞儀評估血小板P-選擇素的活化及血小板之vWF 受器被CD42b (GpIbα) 結合的情形,結果顯示兩種製備過程對於血小板造成的活化程度並無明顯不同(p=0.77 和p=0.62)。另以添加促凝劑 (agonists) 產生的血小板凝集試驗也顯示兩種製備過程對於血小板的凝集能力並無顯著不同。綜合以上之結果,在搖盪儲存前一小時的靜置對於恢復血小板的凝集功能並無明顯的差別。
Platelets play a pivotal role in hemostasis and thrombosis. It induces vascular retraction and clot formation through platelets activation and signal transmission, which promote ligands expressing on the surface of platelets, such as glycoproteins and P-selectin. Some surface glycoproteins gatherd to form complexes after activation. It bound to extracellular receptors such as collagen and thrombin to induce aggregation, which could also be induced by releasing agonists, such as arachidonic acid, adenosine diphosphate, epinephrine, serotonin and fibrinogen, to active the nearby platelets. The standard process of plateletapheresis in the Blood Center was to hold the platelets in still for one hour before stored on a vibrator. The process of holding platelets still for one hour before storage was omitted in some hospitals. It was not clear whether to omit the process has any effect on the quality of platelets. The expressions of P-selectin and vWF receptor, CD42b (Gp Ibα) on platelets were analyzed by flow cytometry in this study. No significant differences (p= 0.77 and p= 0.62, respectively) were found. Similar results were obtained when functions of platelets were evaluated by agonists. It was concluded that leaving the platelets in room temperature for one hour to recover before keeping it on a vibrator would not enhance the functions of platelets aggregation significantly.
中文摘要.........................................................................i
英文摘要.........................................................................ii
英文縮寫表.....................................................................iii
研究目的.........................................................................iv
壹、緒論.............................................................................1
一 、血小板的形成與分解................................................1
二 、血小板的結構............................................................3
三 、血小板的活化與凝血機轉........................................4
四 、血小板在免疫系統中的角色....................................8
五 、血小板相關的疾病..................................................10
六 、血小板分離術 (plateletapheresis ; PH)..............10
貳 、材料方法..................................................................13
一 、設備材料..................................................................13
二 、檢體收集..................................................................13
三 、血小板表面蛋白分析..............................................14
四 、血小板凝集試驗......................................................16
五 、統計分析..................................................................17
參 、結果..........................................................................18
一 、不同血小板分離過程對血小板活化影響..............18
二 、血小板凝集能力之分析..........................................19
三 、不同血小板分離術對血小板凝集能力的影響......20
四 、長期捐贈者與短期捐贈者的血小板功能之比較..20
肆 、討論..........................................................................22
伍 、參考文獻..................................................................27
表目錄...............................................................................33
表一 不同的血小板製備過程對於CD62p (P-selectin) 及CD42b (GpIbα)表現的影響...........................................33
表二 Epinephrine 誘發之血小板凝集與其他促凝劑間之關係..................................................................................34
表三 不同的血小板製備過程對於各種促凝劑凝集能力的影響..................................................................................35
表四 捐贈者之基本資料及捐贈期間長短對血小板凝集能力之影響..........................................................................36
圖目錄..............................................................................37
圖一、不同的血小板製備過程對於CD62p (P-selectin) 表現的情形......................................................................37
圖二、不同的血小板製備過程對於CD42b (GpIbα) 表現的情形..............................................................................38
圖三、花生四烯酸 (Arachidonic acid) 誘發血小板凝集反應圖...................................................................................39
圖四、二磷酸腺苷 (adenosine diphosphate) 誘發血小板凝集反應圖...................................................................40
圖五、膠原 (Collagen) 誘發血小板凝集反應圖..........41
圖六、腎上腺素 (Epinephrine) 誘發血小板凝集反應圖.......................................................................................42
圖七、花生四烯酸 (Arachidonic acid) 之血小板凝集反應分佈圖...............................................................................43
圖八、二磷酸腺苷 (adenosine diphosphate) 之血小板凝集反應分佈圖................................................................44
圖九、膠原 (Collagen) 之血小板凝集反應分佈圖.......45
圖十、腎上腺素 (Epinephrine) 之血小板凝集反應分佈圖........................................................................................46
圖十一、血小板捐贈者平均的凝集情形........................47
圖十二、不同的血小板製備過程對於血小板凝集能力的影響....................................................................................48
圖十三、長期血小板捐贈者與短期血小板捐贈者的血小板凝集功能........................................................................49
附錄 實驗原始資料...........................................................50
附錄一、血小板對於CD62p (P-selectin) 及CD42b (GpIbα) 表現圖..................................................................50
附錄二、血小板對於各種促凝劑的凝集能力圖............80
附錄三、全體血小板捐贈者的新鮮血小板凝集能力圖......................................................................................110
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