碳六十化合物目前已被證實為一有效的氧自由基 (reactive oxygen species, ROS) 的清除劑，本實驗設計的主要研究目的為測試含有氫氧官能基 (-OH groups) 的碳六十化合物polyhydroxylated fullerene derivate C60(OH)9±2 對於RAW 264.7 細胞培養株及失血之後予以再灌流 (ischemia-reperfusion, IR) 的游離肺 (isolated lung) 中所遭受的氧化性傷害 (oxidative stress) 是否具有保護性的作用。
我們在實驗中使用sodium nitroprusside (SNP) 及H2O2分別提高在RAW 264.7細胞培養液中的一氧化氮 (nitric oxide, NO) 及氧化物 (oxidants)。此外，在游離肺進行失血之後予以再灌流的步驟中，我們的觀察目標共分為3個時期：基線時期 (baseline) ，缺血時期 (ischemia)，以及再灌流時期 (reperfusion)。
實驗結果發現，在RAW 264.7 細胞，SNP (1 mM) 或H2O2 (400 mM) 導致細胞存活率下降至90﹪，而隨著 C60(OH)9±2 加入的濃度越高，細胞存活率有明顯提升的現象，與濃度是正相關的關係。另一方面，經由SNP或H2O2 所引發的ROS增加，也能經由C60(OH)9±2 的加入而減少。此外，經由SNP或H2O2 所導致的粒腺體膜電位 (mitochondria membrane potential) 下降的現象也能被 C60(OH)9±2 逆轉。然而，高濃度的 C60(OH)9±2 (1, 1.5 mM) 卻會導致細胞死亡。 在游離肺中，我們也發現在IR進行過程中由SNP所引發的肺動脈壓 (pulmonary artery pressure, Pa) 及肺微血管過濾壓 (pulmonary capillary filtration pressure) 之增高，能被 C60(OH)9±2 減低。
經由以上實驗結果，證明了低濃度的碳六十化合物C60(OH)9±2 可以保護RAW 264.7 細胞培養株及失血-再灌流的游離肺所遭受的氧化性傷害。

Fullerene derivatives have often been used as effective scavengers for reactive oxygen species (ROS). This study was designed to test whether a polyhydroxylated fullerene derivative [C60(OH)9±2] protects against oxidative stress in cultured RAW 264.7 cells and ischemia-reperfused (IR) lungs. We used sodium nitroprusside (SNP) and H2O2 to elevate nitric oxide and oxidants, respectively, in the culture medium of RAW 264.7 cells. In addition, the experimental procedure for the induction of IR-induced injury of isolated rat lungs was divided into three periods: baseline, ischemia and reperfusion. In RAW 264.7 cells, SNP (1 mM) or H2O2 (400 mM) caused a marked (90%) decrease in cell viability, and this decrease was dose-dependently reversed by pretreatment with C60(OH)9±2 (10 to 50 mM). The increase in ROS production induced by SNP or H2O2 was significantly suppressed by C60(OH)9±2. Also, the decrease in mitochondrial membrane potential induced by SNP or H2O2 was significantly reversed by C60(OH)9±2. However, high concentration of C60(OH)9±2 (1 and 1.5 mM) induced cell death (apoptosis or necrosis). In the isolated rat lung, the increases in pulmonary artery pressure and capillary filtration pressure induced by SNP during IR was significantly reversed by C60(OH)9±2 (10 mg/kg). These results indicate that the polyhydroxylated fullerene derivative C60(OH)9±2 in low concentration can protect against oxidative stress in RAW 264.7 cells and IR lungs.

目錄
圖次…………………………………………………………….…….2
中文摘要……………………………………………………………..9
英文摘要……………………………………………………….…….11
前言…………………………………………………………………..13
材料與方法…………………………………………………………..24
實驗結果……………………………………………………………...31
討論…………………………………………………………………...40
結論…………………………………………………………………...50
參考文獻……………………………………………………………...51
附圖…………………………………………………………………...64

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