- 學位論文
天然抗氧化萃取物Raffinee對缺血性中風之效益探討
Study of the effects of the natural antioxidant extract-Raffinee in ischemic stroke
摘要
急性中風 (Acute stroke) 的成因，是由於長期壓力或高脂肪飲食等各種因素所造成，無論在美國、台灣，以及其他國家，已被認為是全球最值得關注的文明病。急性中風又分成出血性中風 (Hemorrhagic stroke) 及缺血性中風 (Ischemic stroke) 。其發生比率各為15%及85%，其中又以男性成年人發生率較高。缺血性中風是由於腦部血管長期受到血栓阻塞，而氧氣及養分無法進入腦部，造成特定區域神經細胞的缺氧及壞死所導致，但目前臨床上對於缺血性中風的治療，只有在黃金三小時內治療最為有效，但往往病患送達醫院時，都超過了黃金三小時，因此，對於超過黃金三小時後的處理及治療藥物，目前尚未明瞭。 缺血性腦中風的傷害，大部分是因為大量的活性氧物種所造成的，而先前有文獻指出，抗氧化劑可延緩缺血性腦中風的傷害。本實驗所使用的藥物-Raffinee，在先前文獻中指出，可治療脊髓損傷，也被應用在臨床上。Raffinee是一種由新鮮的豆子及穀類所萃取出來的一種抗氧化的成分，它對於腦脊髓疾病、糖尿病及相關氧化壓力疾病都有很好的保護作用。基於上述證實，在此將探討Raffinee對於缺血性中風所造成的腦損傷是否有保護作用。本實驗室以8-10週大的雄性C57BL/6J小鼠作為研究動物，再以缺血性中風誘導手術使其右中大腦動脈阻塞，再於每日一次Raffinee的餵食，後續以腦切片、Western Blotting、Zymography等實驗技術分析當動物處於缺血性中風的傷害下，所造成的氧化壓力、發炎、細胞凋亡等路徑相關蛋白的表現情形。由實驗結果發現，有餵食Raffinee的組別，不管是血清中發炎因子、氧化壓力相關指標和控制組比起來，都有明顯的降低，也許是Raffinee是一種抗氧化劑，可透過清除在缺血性中風下，多餘的活性氧物種 (Reactive Oxygen Species,ROS) 來降低MMP2、MMP9的切割活化、CRP和IL-1β的表現，而降低腦部細胞受到缺血再灌注的損傷。因此，Raffinee在未來也許可以提供在治療與預防缺血性中風的另一種選擇。
並列摘要
Acute stroke, which is the most popular diseases in whole the world, including America, Japan, Taiwan and other nations. The results of acute stroke is due to stay in long tern stress or intake too much high fat diet. Acute stroke is also classified with Hemorrhagic stroke and Ischemic stroke. The ratio of these are 15% and 85%, male adult’s occur ratio is higher than another. Due to cerebrovascular is suffered from thromboxane occluding, then oxygen and nutrition can’t access to brain and lead to neuron cells which are in the special zone are hypoxia and necrosis. Recently, clinical has proved that ischemic stroke’s treatment has the best effect of time is in golden three hours, but when stroke patients were arrived at emergency room, the golden three hours had past. Therefore, the drug which could treat stroke patients after golden three hours is unknow. The reasons of harm in ischemic stroke is that the most reactive oxygen species (ROS) aggress brain tissues. Recently, some researchs have suggested that antioxidant can ameliorate the harm of brain tissue in ischemic stroke. So we’ll use the drug- Raffinee to study our experiment. Raffinee is extracted from bean and bran, it has strong antioxidant ability. Raffinee has the best protective effect in spinal cord disease, diabetes and some relative oxidative stress disease. According to above result, we’ll study that whether Raffinee could protect brain tissue injury or not. We use 8-10 weeks old male C57BL/6J mice as ischemic stroke animal model. Then we do some MCAO surgery to mimic ischemic stroke disease in animal model. Then we give some Raffinee to C57BL/6J mice once a day. After sacrificing, we use some experiment technique such as brain slice, Western blotting, Zymography and so on to analysis when mice’s brain is suffered from ischemic stroke damage like oxidative stress, inflammation, cell necrosis and apoptosis relative signaling pathway. These results reveal that the group fed Raffinee whether serum inflammatory factor, oxidative stress relative marker are lower than another group. Perhaps, Raffinee could through clean more ROS to decrease MMP2, MMP9’s cleavage and activation, CRP, IL-1β’s expression and decrease brain cell’s injury. Therefore, Raffinee will provide treatment and prevent in ischemic stroke in the future.