16338.irgarol1051通過破壞線粒體的結(jié)構(gòu)和功能而誘發(fā)細胞凋亡

1、IrgarolIrgarolIrgarolIrgarol1051105110511051通過破壞線粒體的結(jié)構(gòu)和功能而誘發(fā)細通過破壞線粒體的結(jié)構(gòu)和功能而誘發(fā)細胞凋亡胞凋亡作者：王麗指導(dǎo)教師：劉文華教授摘要本文通過Irgarol1051對HepG2細胞進行處理后發(fā)現(xiàn)，細胞線粒體的產(chǎn)生瞬時ROS，進而破壞線粒體脊的結(jié)構(gòu)和其功能，使其膜電位降低，促進線粒體釋放細胞色素C，以至細胞凋亡程序啟動而影響細胞的正常代謝活動，使細胞的生長受到抑制，進而誘

2、發(fā)細胞凋亡。通過進一步的體外線粒體暴露實驗得出，Irgarol1051對線粒體作用的深層機制是干擾電子從細胞色素氧化酶進行傳遞進程，同時抑制了復(fù)合物Ⅲ的活性而最終抑制ATP的合成，而其引發(fā)的線粒體功能紊亂最終也誘導(dǎo)了細胞的死亡。同時Irgarol1051和Cu2的協(xié)同暴露實驗發(fā)現(xiàn)，，而使細胞的生長受到抑制，進而誘發(fā)細胞凋亡；同時，HepG2細胞切片的電鏡觀察顯示，可以提高促凋亡基因BAX的表達而使細胞凋亡，同時表現(xiàn)出一定的神經(jīng)毒性。關(guān)鍵

3、詞：Irgarol1051；細胞凋亡；線粒體；BAX；細胞色素C目錄中文摘要...........................................................................................................................Ⅰ英文摘要......................................................

4、.....................................................................Ⅱ目錄...................................................................................................................................Ⅲ第一章緒論...........

5、..........................................................................................................71.1研究背景..........................................................................................................

6、....71.1.1Irgarol1051的檢測及應(yīng)用.......................................................................71.1.2Irgarol毒性研究的現(xiàn)狀..........................................................................121.2細胞凋亡...................

7、.........................................................................................131.3氧化應(yīng)激與細胞損傷.......................................................................................141.4線粒體與細胞凋亡的關(guān)系..............

8、.................................................................161.5電子傳遞鏈........................................................................................................171.6NO的神經(jīng)毒性作用...........................

9、...............................................................181.7研究目的............................................................................................................19第二章實驗材料...............................

10、...........................................................................19第三章實驗方法..........................................................................................................203.1Irgarol誘導(dǎo)HepG2細胞凋亡及機制研究..

11、....................................................203.1.1細胞活力檢測..........................................................................................203.1.2AnnexinVFITCPI雙染法.........................................

12、...........................213.1.3細胞內(nèi)ROS含量檢測...........................................................................223.1.4線粒體膜電位檢測.................................................................................223.1

13、.5細胞內(nèi)鈣離子濃度.................................................................................233.1.6線粒體呼吸爆發(fā).....................................................................................233.1.7Caspase3酶活力檢測..........

14、....................................................................233.1.8Caspas9活力檢測.................................................................................243.1.9細胞核DAPI熒光染色...................................

15、.......................................243.1.10細胞內(nèi)CytochromeC檢測.................................................................253.1.11細胞組織電鏡切片............................................................................

16、...253.2體外線粒體Irgarol暴露研究其對電子傳遞的影響.....................................263.2.1線粒體的提取..........................................................................................263.2.2線粒體蛋白質(zhì)定量................................

17、.................................................263.2.3線粒體完整性測定（RCR測定）.......................................................273.2.4線粒體體外ROS測定...........................................................................27

18、3.2.5線粒體體外膜電位測定.........................................................................273.2.6線粒體呼吸鏈復(fù)合物V活性測定.......................................................283.3Irgarol與Cu2對大鼠神經(jīng)細胞的協(xié)同效應(yīng)............................

19、.......................283.3.1NO熒光檢測............................................................................................283.3.2乙酰膽堿酯酶(AChE)檢測....................................................................283

20、.3.3乙酰膽堿轉(zhuǎn)移酶（ChAT）檢測..........................................................293.3.4抑制凋亡因子Bcl2和促凋亡因子BAX的測定..............................29第四章.實驗結(jié)果..........................................................................

21、...................................294.1Irgarol誘導(dǎo)HepG2細胞凋亡...........................................................................294.1.1Irgarol抑制HepG2細胞生長..............................................................