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研究生:楊承憲
研究生(外文):Cheng-Hsien Yang
論文名稱:香椿萃取物對氧化壓力下HepG2細胞的保護機制
論文名稱(外文):The protective mechanism of Toona sinensis Roem extracts under oxidative stress in HepG2 cells
指導教授:張素瓊張素瓊引用關係
指導教授(外文):Sue-Joan Chang
學位類別:碩士
校院名稱:國立成功大學
系所名稱:生命科學系碩博士班
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:88
中文關鍵詞:細胞凋亡氧化壓力香椿
外文關鍵詞:Toona sinensisapoptosisoxidative stress
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  • 被引用被引用:1
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香椿(Toona sinensis Roem)為東南亞常見之食用及藥用植物,具有抗氧化、抗發炎、改善糖尿病等諸多保健功效。近年研究發現,香椿中主要的成分為槲黃素(quercetin)及沒食子酸(gallic acid),具有良好的抗氧化能力。本研究主要探討不同香椿萃取物對過氧化氫(hydrogen peroxide, H2O2)誘發HepG2細胞株氧化傷害的保護機制。將HepG2細胞預先以香椿萃取物(TSL-2、TSL-2P及TSL-6)、quercetin及gallic acid處理,再給予細胞氧化傷害後,分析細胞的存活率,並探討是否造成細胞凋亡及分析Mitogen activated protein kinases(MAPKs)及caspases等訊息傳遞路徑是否參與其中。研究結果發現,H2O2會造成HepG2細胞內ROS顯著上升、降低粒線體膜電位並造成細胞存活率顯著下降,經流式細胞儀及LDH釋放量分析證實H2O2造成的細胞死亡主要為細胞凋亡(apoptosis)而非細胞壞死(necrosis);利用caspases及MAPKs的抑制劑及西方墨點法(western blotting)分析證實H2O2會促使p-ERK及p-JNK蛋白質表現量上升、增加caspase-9及caspase-3的活化態、促使pro-apoptotic蛋白質Bax、Smac/DIABLO表現量上升並抑制anti-apoptotic蛋白質Bcl2的表現。細胞預先以quercetin、TSL-2P及TSL-6處理能有效降低胞內ROS、維持粒線體膜電位、降低細胞凋亡的比率、抑制caspase-9及caspase-3的活化、降低p-ERK、p-JNK、Smac及Bax的蛋白質表現量,並增加Bcl2的蛋白質表現;細胞預先以gallic acid處理可稍加抑制caspase-9的活化及增加Bcl2的蛋白質表現,但是無法降低由H2O2誘使的細胞凋亡;給予細胞先以TSL-2處理可稍加抑制caspase-9的活化、增加Bcl2的蛋白質表現及降低Bax的表現量,但是無法降低由H2O2誘使的細胞凋亡。綜合以上結果,預先處理細胞槲黃素和香椿萃取物(TSL-2P and TSL-6)處理可以降低胞內ROS,並可透過MAPKs訊息傳遞路徑和caspases蛋白質路徑抑制由H2O2誘使的細胞凋亡。本研究結論,香椿可以減緩由氧化傷害誘發的細胞凋亡,未來可更加推廣其食用及藥用價值並積極開發為抗氧化傷害保健食品。
Toona sinensis Roem (TS) is a traditional Chinese health food reported to exhibit many beneficial physiological functions. Hydrogen peroxide (H2O2)induced powerful oxidative stress in cells and led to apoptosis. In this study we focused on the protective mechanisms of extracts from TSL(TS Leaves) (TSL-2, TSL-2P and TSL-6 ). HepG2 cells(human liver carcinoma cells) were pre-treated with TSL-2, TSL-2P, TSL-6, gallic acid and quercetin(two major active compounds of TS)and then treated with H2O2. Cell viability was measuredd by MTT assay and showed that H2O2 induced cell death recovered by prereatment of TSL-2P, TSL-6 and quercetin. Reactive oxygen species(ROS)production and apoptotic cells measured by flow cytometry were significantly higher in H2O2 group than that in TSL-2P, TSL-6 and quercetin groups. Activated caspase-3 and caspase-9 were significantly higher in H2O2 group than that in TSL-2P, TSL-6 and quercetin groups by western blotting. Cell death induced by H2O2 was reversed by MAPKs inhibitors, and TSL-6. Protein expression of p-ERK and p-JNK was significantly higher in H2O2 group than that in TSL-2P, TSL-6 and quercetin groups. In addition, pro-apoptotic proteins Bax and Smac were significantly higher in H2O2 group than those in TSL-2P, TSL-6 and quercetin groups. However, anti-apoptotic protein Bcl2 was significantly lower in H2O2 group. In conclusion, TSL-2P, TSL-6 and quercetin may recover the H2O2 induced apoptosis via MAPKs and csapases pathway.
中文摘要---------------------------------------------- I
英文摘要---------------------------------------------- III
誌謝---------------------------------------------------IV
目錄-------------------------------------------------- V
第一章:前言------------------------------------------ 1
第二章:文獻探討-------------------------------------- 3
第三章:材料與方法------------------------------------ 14
第四章:結果-------------------------------------------25
第五章:討論-------------------------------------------34
第六章:參考文獻---------------------------------------46
圖---------------------------------------------------- 61
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