姜黄素诱导的DNA损伤和细胞凋亡及其机制的研究-营养与食品卫生学专业论文.docxVIP

复旦大学硕士学位论文 复旦大学硕士学位论文 英文摘要 Abstract 【objeetive]：Curcumin(diferuloylmethane，CUR)，a polyphenol derived from the plant Curcuma longa，has been widely consumed in food，drug and cosmetic．Like some well．known anfioxidants．the effect of curcumin alSO iS“a two．edged sword’L --Mthough curcumin is a naturally occurring antioxidant，it exhibits prooxidant properties under certain conditions．To study the oxidative damage induced by curcumin，tlle reaction conditions，the dose-response relationship and time—effect relationship，and its possible mechanism． IMethods]：(1)Curcumin caused the damage ofcell DNA：Combined single cell gel electrophoresis assay with cytoactive determination．DNA damages and repairs of CHL cell and SMMC7721 cell were respectively detected，which was caused by 0～1 00Ⅲnol／L curcumin with，or non-CU2+．(2)Relationships between apoptosis and oxidative stress：Curcumin treated the CHL cells and也ell cellular morphological changes were observed witll the microscope．Analysis ofaccounts for cell cycle and apoptotic percent the percent of apoptosis was measured、Ⅳitll flow cytometry．The percent of apoptosis and cell membranes damages were detected、vith EB／AO double staining method．Intra·cellular ROS was also measured with flow cytometry by DCFH-DA as Molecular Probes．(3)Cureumin induced the oxidative damage of plasmid pBR322 DNA：Firstly．eurcuminⅥ，as incubated with plasmid pBR322DNA for 0--4 hours under 37℃circumstance that coexisted with different levels ofCu” (0～2001amol／L)or antioxidants．Then the electropherogram was scanned after agarose gel electrophoresis．Finally，the pixel values ofvarious configurations for plasmid DNA were measured and percent of supercoil DNA was calculated． [Resultsl：(1)④After O-lOOpmol／L curcumin without CU2+stimulated CHL cells for 2 hours，survival rates of CHL cells were all above SOpercent．However, 100Bmol／L curcumin caused the survival rate of SMMC7721 eell to drop below 80 percent．Adding 1 00 Ixmol／L Cu”．the survival rates of CHL cell