最近有研究發現，將E.coli lipopolysaccharide (LPS)注射到大鼠後，一直到個體死亡之間，測量系統動脈壓(SAP)時，會伴隨產生低頻訊號(0-0.8 Hz)的變動。整個低頻訊號的變動可分三期，包括第一期的訊號降低，第二期的訊號增強，第三期的訊號再度降低。而這種低頻訊號有人認為與腦幹有關，甚至認為與腦幹中的rostral ventrolateral medulla (RVLM)有密切關係。而我們想了解在這低頻訊號變動的生理現象中，有哪些差異表現的基因存在，並且這些基因是否有參與腦幹死亡的過程。所以我們利用一種對於找尋差異表現基因非常有效的方法，suppression subtractive hybridization(SSH)(抑制性扣減雜交)，並且建立基因庫，來找出在RVLM第一期與第二期之間的差異表現基因。在我們從基因庫初步篩選的基因，經過資料庫比對之後，到目前為止，我們發現一些在RVLM第一期與第二期之間，有差異表現的基因，這些基因有些是正向調控表現，有些則是負向調控表現。所以這些基因的差異表現，可能參與大鼠腦幹死亡過程中的分子調控機制。

Recent studies have discovered that LPS-treated Sprague-Dawley rats induced a reduction (phase I), followed by an augmentation (phase II), and decrease again (phase III) in the power density of the vasomotor component (0-0.8 Hz) in systemic arterial pressure (SAP). It was reported that the vasomotor components were related to the brain stem, even closely related to the rostral ventrolateral medulla (RVLM). But the molecular mechanism involved in the death progression of rat brain stem is mostly unknown. We used suppression subtractive hybridization (SSH) and library construction to find differentially expressed genes between phase I and phase II of LPS-treated RVLM. At present, we have found some genes that are differentially expressed between phase I and phase II of LPS-treated RVLM. Some genes are up-regulation expression and others are down-regulation expression. Thus, these genes may be involved in the molecular mechanism of the death progression in the rat brain stem.

中文摘要------------------------------------------------------------------p.1
英文摘要------------------------------------------------------------------p.2
緒 論------------------------------------------------------------------p.3
腦幹死亡與動脈壓頻譜訊號----------------------------------------------p.3
Lipopolysaccharide (LPS) ------------------------------------------------p.4
Subtractive hybridization and subtractive cDNA library -------------p.7
Suppression subtractive hybridization (SSH) -------------------------p.10
實驗目的------------------------------------------------------------------p.13
實驗材料與方法------------------------------------------------------------p.14
實驗結果------------------------------------------------------------------p.28
抑制性扣減雜交與基因庫的建立---------------------------------------p.28
由點雜合實驗找到一些差異表現的基因------------------------------p.30
DNA序列與生物資訊資料庫基因比對之結果-----------------------p.32
以RT-PCR 實驗做差異表現基因之初步確認------------------------p.33
討 論-------------------------------------------------------------------p.35
Subtractive hybridization -------------------------------------------------p.35
差異表現基因---------------------------------------------------------------p.36
參考資料-------------------------------------------------------------------p.47
附 圖-------------------------------------------------------------------p.53

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