【作者】 陆姝欢；

【导师】 元英进；

【作者基本信息】 天津大学 ， 制药工程， 2007， 硕士

【摘要】 为了研究正常生长的东北红豆杉细胞和自发凋亡的南方红豆杉细胞磷脂代谢的差异,本课题运用液质联用技术(HPLC/ESI/MSn)从脂质组学(lipidomics)的角度,对两种红豆杉细胞磷脂代谢进行了系统水平的研究。主要研究结果为:(1)建立了以HPLC/ESI/MSn为基础的高通量、高灵敏度及高分辨率的研究红豆杉细胞磷脂的分析方法;(2)发现了红豆杉细胞中至少含有6类(class)84种(species)磷脂分子,以此为基础建立了红豆杉细胞的磷脂图库。同时新发现了红豆杉细胞内的磷脂含有特殊的多不饱和长碳链,如C20:3及C20:4等;(3)测定了东北红豆杉细胞正常生长和南方红豆杉细胞自发凋亡过程中磷脂含量的变化,并采用主成分分析方法(PCA)对主要的77种磷脂的代谢变化进行研究。该方法能明显区分自发凋亡的南方红豆杉细胞与正常生长的红豆杉细胞,确定了PC和PA两类磷脂是主要的生物标志物群,且在第一主成分上呈强负相关,揭示了PC和PA的代谢异常可能在南方红豆杉细胞自发凋亡过程中起关键作用。基于PA是重要的第二信使,参与细胞凋亡的信号转导,激活MAPK等下游路径。推断引起南方红豆杉细胞磷脂变化的原因可能是其细胞中的磷脂酶D被激活、作用于PC,将其降解为PA,最终导致细胞的自发凋亡。为进一步发掘磷脂在南方红豆杉细胞凋亡过程中的作用机制建立了基础。更多还原

【Abstract】 In order to elucidate the difference of phospholipid metabolism in two Taxus species,Taxus cuspidata and Taxus chinensis, we employ the lipidomics strategy to systematically study the phospholipids in the two species by HPLC/ESI/ MSn. The achievements in our study are as follows: (1) We establish a method of high throughput, high sensitivity and high discerning capacity to analyze the phospholipid profiles in Taxus cells. (2) We found that there are over 84 molecular species of 6 different phospholipids classes in Taxus cells, and we establish a Taxus phospholipid database basing on these results. A new discovery in this study was that the phospholipids in Taxus cells contain specific unsaturated long carbon chains, such as C20:3 and C20:4. (3) We measured the changes of phospholipid contents in normally cultured Taxus cuspidata and the automatically apoptotic Taxus chinensis. Through principal analysis of the main 77 species of phospholipids, the normally cultured Taxus cuspidata and the automatically apoptotic Taxus chinensis could be evidently distinguished. And the two negative correlating species on the first principal component-PC and PA mainly contributing to the distinguishment, and thus they may be the important biomarkers and their abnormal metabolism may play key roles in the automatic apoptosis of Taxus chinensis.In conlusion, we presume that the changes of phospholipid in Taxus chinensis are due to the activation of PLD(phospholipase D) and its subsequent degeneration of PC to PA, which finally lead to the automatic apoptosis. Our study establishes the foundation for further exploration of the phospholipid’s roles on Taxus chinensis’apoptosis.更多还原