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研究生:李宜螢
研究生(外文):Lee, Yi-Ying
論文名稱:紫色甘藷萃取物改善胰臟ß細胞氧化壓力誘發之凋亡現象
論文名稱(外文):Purple Sweet Potato Extracts Improve Oxidative Stress-induced Pancreatic β-cells Apoptosis
指導教授:林佳育
指導教授(外文):Lin, Chia-Yu
口試委員:殷梅津鄭統隆林佳育
口試委員(外文):Yin, Mei-ChinCheng, Tung-LungLin, Chia-Yu
口試日期:2012-07-23
學位類別:碩士
校院名稱:亞洲大學
系所名稱:保健營養生技學系碩士班
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:中文
論文頁數:56
中文關鍵詞:胰臟β細胞紫色甘藷氧化壓力
外文關鍵詞:Pancreatic β-cellspurple sweet potatooxidative stress
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胰臟β細胞相較於其他器官中之細胞,具有相對較少之抗氧化酵素,因而使β細胞更容易受到氧化損傷。過往文獻中已暗示紫色甘藷purple sweet potatos(PSP)具有抗氧化、抗發炎,以及控制血糖之作用,意味著PSP對糖尿病的潛在好處,因此本研究探討PSP萃取物改善胰臟β細胞因氧化壓力而誘發之凋亡現象。將INS-1細胞予以毒性(H2O2或STZ)或PSP不同濃度及不同作用時間處理,以MTT、Sub G1期及西方墨點法偵測細胞存活情況,並使用DPPH清除、總酚含量、亞鐵離子螯合、還原力、細胞抗氧化酵素活性(SOD、GPx及Catalase)與ROS清除能力等試驗進行與抗氧化相關之測試,此外,利用JC-1染色、ATP試驗及胰島素分泌實驗偵測胰臟β細胞功能。結果顯示,PSP能顯著回復由氧化壓力所誘導之細胞死亡,且抑制caspase-3等凋亡相關蛋白的表現,另外,PSP具50%以上清除因氧化壓力誘導而產生ROS的能力,並有效提升細胞內抗氧化酵素的活性,於β細胞之功能狀態結果顯示,PSP可緩解粒線體膜電位去極化現象、使因氧化壓力而降低之ATP產量回升,且回復β細胞胰島素分泌功能。整體而言,本研究結果顯示PSP可改善胰臟β細胞因氧化壓力而誘發之凋亡現象及回復胰島素分泌功能,此結果可能部分與PSP之自由基清除能力及維持粒線體正常功能有關。
Pancreatic β-cells are relatively low in the expression of antioxidant enzymes, which render β-cells more susceptible to oxidative damage. It has been implied that purple sweet potatos (PSP) have anti-oxidative and anti-inflammatory effects, as well as may improve blood glucose levels, suggesting the potential benefits of PSP on diabetes. Therefore, the purpose of this study was to investigate the anti-apoptotic effects of PSP on pancreatic β-cells. INS-1 cells were treated with PSP and/or H2O2/STZ for variuos concentrations and time periods. Cell survival was detected by MTT, Sub G1 phase and western. The antioxidative effect of the PSP was determined by DPPH radical-scavenging activity, total phenolic determination, ferrous ion chelating activity, reducing power, Superoxide Dismutase (SOD) Activity, Glutathione Peroxidase (GPx) Activity, Catalase Activity and reactive oxygen species (ROS) assay. In addition, JC1 analysis and ATP assay, as well as insulin secretion were conducted to measure intracellular functions. Treatment of H2O2/STZ causes more than 50% of cell death, which is recovered by PSP treatment significantly. This improved cell survival by PSP could be due to the reduced caspase-3 expression. Moreover, PSP might act as an atioxidant, which inhibits more than 50% of the oxidative stress-induced intracellular ROS generation. Cellular ATP content and mitochondria membrane potential are also alleviated in the presence of PSP after oxidative stress. While insulin secretion after H2O2/STZ treatment is about one third of that of control, PSP dramatically recovers it. In summary, these findings indicate that PSP protects pancreatic β-cells from oxidative stress-induced apoptosis and improves insulin secretion. This enhanced cell survival could be partially due to the ability of PSP to scavenge free radical and maintain mitochondria function.
目 錄
中 文 摘 要 i
Abstract ii
誌 謝 iii
目 錄 iv
圖目錄 vi
表目錄 vii
第一章 前言與研究目的 1
第二章 文獻探討 2
第一節、胰臟Β細胞之功能與特性 2
第二節、胰臟Β細胞的死亡 2
第三節、氧化壓力對胰臟Β細胞之影響 4
1.粒線體損傷 5
2.內質網損傷 6
第四節、研究材料介紹 7
第三章 材料與方法 9
樣品製備 13
DPPH清除率 13
亞鐵離子螯合 13
還原力試驗 14
多酚總含量測定 14
細胞培養 15
MTT細胞存活率 15
偵測SUB G1期實驗 15
細胞內生性活性氧REACTIVE OXYGEN SPECIES (ROS)生成含量 16
細胞抗氧化酵素SUPEROXIDE DISMUTASE (SOD)活性 16
細胞抗氧化酵素CATALASE活性 17
細胞抗氧化酵素GLUTATHIONE PEROXIDASE (GPX)活性 17
細胞粒線體膜電位變化 18
細胞ATP含量 18
西方墨點法蛋白轉漬 19
細胞胰島素分泌 20
統計分析 20
第四章 結果 21
DPPH清除率 21
亞鐵離子螯合 21
還原力試驗 21
多酚總含量測定 22
MTT細胞存活率 22
偵測SUB G1期實驗 23
細胞內生性活性氧REACTIVE OXYGEN SPECIES (ROS)生成含量 24
細胞抗氧化酵素SUPEROXIDE DISMUTASE (SOD)活性 24
細胞抗氧化酵素CATALASE活性 25
細胞抗氧化酵素GLUTATHIONE PEROXIDASE (GPX)活性 25
細胞粒線體膜電位變化 26
細胞ATP含量 27
西方墨點法蛋白轉漬 27
細胞胰島素分泌 29
第五章 討論 30
第六章 結論 37
參考文獻 38
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