臺灣博碩士論文加值系統

　　脾臟全切除加快肝臟再生此一現象已早被注意到，然而何種因素所造成至今依然沒有定論，是肝臟增生因子增加表現亦或抑制因子減少值得吾人探討。本研究以切除大鼠2/3 肝臟以引發肝臟再生之動物模式，術後以增生之全部肝臟重量與大鼠體重之比值相比較，實驗組(切除2/3 肝臟加上脾臟全切除) 於術後之第一天及第二天高於對照組(只切除2/3 肝臟)。以RT-PCR測定肝臟IL-6表現量，實驗組明顯低於對照組，血清學上IL-6濃度於實驗組大為降低。有學者發現來自脾臟TGF-ß1 與再生肝臟中的TGF-ß receptor II，會影響肝再生情形，本研究也可見到相似結果。以RT-PCR測定肝臟HGF (hepatic growth factor) mRNA的表現，在手術後一小時及兩小時實驗組高於對照組；而抑制性因子Activin-a方面，實驗組則於二、四、六小時明顯低於對照組；以Spot-denso 測定每一時間點之相對量， 以HGF與Activin-a之比值相比較，實驗組依然高於對照組，其中HGF在最早之時間點即已有所改變，可能與切除2/3 肝臟再加上脾臟全切除時造成之門脈血流動力學變化有關，但詳細機轉仍須詳加探討。在蛋白質層次上，以西方墨點也證實Activin-a表現量在對照組確實隨著時間點逐漸降低，而實驗組在相對時間點上更較對照組為低，參考過去文獻，TGF-ß1在部分切除肝臟後較手術前為高，然而其接受體卻降低，造成TGF- ß1 在肝臟再生早期“不敏感” (insensitivity)，因此脾臟全切除加快肝臟再生的可能機轉上Activin-a似乎扮演更重要之角色，然而在臨床上仍須更詳盡的實驗數據及詳細機轉支持以茲運用。

　Splenectomy enhanced liver regeneration was noticed for a long time. However, which factor was involved in this phenomenon is inconclusive. In this study, we used partial hepatectomised rat as a animal model to observe the expression of factors in liver regeneration. The ratio of regenerating liver and rat body weight at the time point 1 and 2 days in experimental group (partial hepatectmoy and splenectomy) is greater than that of control group (partial hepatectmy). Expression of RT-PCR of IL-6 in liver tissue of experimental group is lower than that of control group. ELISA for serum IL-6 confirmed above phenomenon. In previous research, TGF-ß 1 released from the spleen exerts a growth inhibitory effect on liver regeneration in rats. Results of this study confirm decreased expression of mRNA of TGF-ß 1 in experimental group than that of control group. It may be due to hemodynamic changes that induced HGF expression at early time point 1 and 2 hours when partial hepatectomy and splenectomy were done simultaneously. Messenger RNA expression of Activin-a in experimental group is lower than that of control group post operation 2、4 and 6 hours. Western blot showed decreased expression of Activin-a with time at control group that is more evident than experimental group at protein level. In previous papers, TGF-ß increased after hepatectomy but hepatocytes become insensitive to it in early phase due to down regulation of receptors on hepatocytes. Activin-a may play a important role for the enhancement of liver regeneration in the early period by splenectomy when TGF-ß signaling did not work. More data and detailed mechanism should be collected to unravel this problem for clinical application.

目錄
中文摘要…………………………………………………Ⅰ
英文摘要…………………………………………………Ⅲ
誌謝 ………………………………………………………Ⅴ
目錄 ………………………………………………………. VII
圖表目錄 …………………………………………………IX第一章、緒……………………………………………… 1
第二章、實驗材料及實驗方法…………………………9
第一節、實驗材料………………………………………9
1. 實驗動物 9
2. 實驗藥品與試劑 9
3. 抗體 11
4. 重要器材 12
第二節、實驗方法…………………………………… 13
1. 動物模式的建立 13 (1)
大鼠肝臟部份切除模式 13
2.形態學評估 14
3以ELISA測定肝門靜脈及下腔靜脈血清細胞激素IL-6濃度 14
4.反轉錄酶及聚合酶連鎖反應 15
5.西方墨點法 17
6.統計分析 17
第三章、結果………………………………………18
1.脾臟切除對於再生肝臟重量及細胞增生指標之效應 18
2.脾臟切除後，增生促進因子訊息RNA之表現變化 18
3.脾臟切除後，其他增生促進因子訊息RNA之表現變化 19
4.脾臟切除後，抑制性因子訊息RNA之表現變化 19
5.肝臟部分切除後再加上脾臟全切除，抑制性因子Activin-a 於肝臟組織之蛋白質表現表現較只有切除部分肝臟之表現較為減弱 20
第四章、討論……………………………………….21
第五章、參考文獻…………………………………27
第六章、圖表………………………………………31

圖表目錄
圖一、實驗摘要圖
圖二、大鼠切除2/3肝臟及脾臟手術圖
表一、再生肝重及鼠體重之比值
圖三、再生肝重及鼠體重之比值以柱狀圖表示
圖四、Mitotic figure 有絲分裂相
圖五、Immunohistochemistry for cell proliferation.
圖六、RT-PCR for Il-6 mRNA
圖七、以ELISA測定血清IL-6 濃度
圖八、RT-PCR for PAI-1 mRNA
圖九、RT-PCR for HGF mRNA
圖十、RT-PCR for TGF-beta 1 mRNA
圖十一、RT-PCR for Activin-a mRNA
圖十二、Rratio of RT-PCR of HGF and Activin-a
圖十三、Western blot of Activin-a in Liver
圖十四、脾臟切除使肝臟Activin-a表現減少
圖十五、再生肝臟重量與體重之比值與肝臟組織Activin-a表現呈負相關趨勢變化。
圖十六、脾臟切除使早期再生肝臟Activin-a蛋白表現減少。

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