臺灣博碩士論文加值系統

腹主動脈瘤在年齡高於60歲的族群發生率是百分之8，且當其破裂時會導致很高的死亡率。腹主動脈瘤的特徵主要是在動脈管壁中層的細胞外間質受到破壞及血管平滑肌細胞的缺失。此外有報導指出c-Jun N-terminal Kinase在形成腹主動脈瘤的動物模式中扮演著重要的角色。先前的文獻中提到血管收縮素二會透過p53/p21此路徑導致血管平滑肌細胞的提前老化。所以我們在此假設c-Jun N-terminal kinase的活化仲介了血管平滑肌細胞的老化及缺失在腹主動脈瘤的形成過程中。我們利用血管收縮素二處理人類血管平滑肌細胞二十四小時及四十八小時後，可以誘發血管平滑肌細胞的提前老化藉由偵測在酸鹼值等於6.0的老化相關β-galactosidase活性。我們也在20 周歲Apo E 缺失老鼠的升主動脈基部的動脈硬化處發現有老化的細胞出現，但是在21周的B6老鼠中並沒有偵測到老化的細胞。我們檢測人類腹主動脈瘤檢體的JNK表現與磷酸化的程度，並以冠狀動脈繞道手術時在升主動脈打洞所取得的檢體當作對照組。發現JNK2活化程度在腹主動脈瘤中比冠狀動脈繞道手術時在升主動脈打洞所取得的檢體中有顯著的增加，但JNK2的表現在兩者檢體間並無差異。相反的，JNK1的活化很低且偵測不到其表現量在兩者檢體中。有趣地，在體外培養的人類動脈血管平滑肌細胞的實驗中，血管收縮素二刺激細胞的JNK1活化在五分鐘後達到最大值且在六十分鐘後回到基礎值但是並沒有偵測到JNK2的活化。然而，血管收縮素二處理血管平滑肌細胞直到四十八小時後，JNK1的活化及表現量似乎有隨血管收縮素二刺激時間增長而上升的趨勢但對照主相比並無達到顯著差異。此外並無JNK2活化被偵測到且其表現量也無明顯的改變在長時間血管收縮素二處理下。利用血管收縮素二處理血管平滑肌細胞四十八小時後，p21Cip1 的表現量並無顯著的增加。血管收縮素二刺激平滑肌細胞2分鐘候可以誘發過氧化氫的產生持續到5分鐘，且隨著時間過氧化氫的產生有減少的趨勢，然而過氧化氫在60分鐘刺激時產量再度增加。更進一步發現，將血管平滑肌細胞處理JNK的抑制劑SP600125，可以阻斷因為血管收縮素二所誘發的血管平滑肌細胞老化的現象。這些結果可能可以說明JNK的活化,可能是JNK1活化,參與在血管收縮素二所誘發平滑肌細胞老化。

Abdominal aortic aneurysm (AAA) is present in 8% of the population over 60 years of age and exhibits a high mortality rate when rupture. AAA is characterized by extracellular matrix destruction and depletion of vascular smooth muscle cells (VSMC) in the tunica media. Angiotensin II (Ang II) was shown to induce AAA formation in animal models and c-Jun N-terminal kinase (JNK) was shown to play a critical role. A previous study suggested that Ang II induces premature senescence of VSMCs via a p53/p21-dependent pathway. We hypothesized that JNK activation mediates VSMC senescence and loss during AAA formation. Ang II treatment for 24 or 48 h induced cellular senescence of VSMC assessed by senescence-associated β-galactosidase (SA β-gal) activity. SA β-gal-postive cells were detected in atherosclerotic lesions of the aortic root in 20 weeks-old ApoE-/- mice but not in the aortic root of 21 weeks-old B6 mice. JNK phosphorylation and expression levels were examined in specimens of AAA and punctured ascending aorta from coronary artery bypass graft (CABG) surgery as control. JNK2 phosphorylation levels in AAA specimens were markedly higher than those in CABG specimens. In contrast, JNK1 expression was not detectable in those specimens with low phosphorylation levels. Interestingly, in cultured human aortic SMCs (HASMCs), Ang II stimulated JNK1, but not JNK2, phosphorylation that peaked at 5 min and returned to basal levels at 60 min. Long-term Ang II treatment up to 48 h tended to increase JNK1 phosphorylation and expression but did not reach significant difference. Neither phosphorylation nor changes in expression of JNK2 was detected in HASMCs following long-term Ang II-treatment. Moreover, JNK inhibition by treating SP600125, a selective JNK inhibitor, inhibited Ang II-induced cellular senescence of HASMCs. The expression of p21Cip1 did not increase significantly after Ang II stimulation. Upon Ang II stimulation, hydrogen peroxide (H2O2) production in HASMCs increased at 2 min and 5 min decreased to near basal levels, and then increased again at 60 min. These results suggested that JNK activation, possibly JNK1, may play a role in Ang II-induced cellular senescence of VSMC.

Chinese Abstract………………………………………………Ⅱ
Abstract…………………………………………………………Ⅳ
Introduction……………………………………………………1
Materials and Methods……………………………………… 13
Results………………………………………………………… 32
Discussion………………………………………………………37
References………………………………………………………44
Figures………………………………………………………… 50
Appendix…………………………………………………………59

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