不同类型细胞对羰基应激的响应差异
发布时间：2018-09-06 19:20
【摘要】： 衰老是遗传因素和环境因素共同作用的结果。根据羰基应激衰老学说的观点,在氧自由基引起的脂质过氧化和非酶糖基化反应过程中,会生成具有反应活性的共同中间产物——活性羰基类物质,它们能与生物体内的几乎所有生物大分子发生羰-氨交联,这一过程是机体内衰老损变的核心过程,甚至是机体衰老改变的生化本质。 本学位论文以分别代表两个极端的正常细胞(即体外生存能力很弱的红细胞和长期生活在体外的酵母)为实验模型,探讨它们对羰基应激的响应差异。 一、血液在储存过程中,血液粘度会随着储存时间的延长逐渐上升,然而其具体生化机制仍不明了。为了探索这种血液粘度升高的机制,本文探索了血液储存过程中硫醇、TBARS水平、红细胞膜蛋白羰基化程度以及蛋白质荧光强度的变化。结果显示:伴随着血液粘度的升高,硫醇水平与蛋白质荧光强度逐渐降低,而TBARS水平与膜蛋白羰基化程度逐渐增加。这些结果表明:活性羰基类物质很容易攻击蛋白质的氨基和巯基,羰基应激诱导的红细胞膜蛋白羰基化可能是血液储存过程中粘度上升的重要机制。二、为了探索酵母细胞对羰基应激的响应,本文用不同浓度的MDA溶液处理酵母细胞,培养过夜后用紫外分光光度计在600 nm处测定培养过夜后酵母的光密度,同时用PI荧光染色法(Ex530/Em590)测定经MDA处理后的酵母细胞的活力。结果显示:即使在高浓度MDA的作用下,酵母细胞生长和活力均未受到显著抑制。这表明:酵母细胞抗羰基应激的能力很强,但这种抗性的具体机制仍需做进一步的实验探讨。 综上所述,血液中的红细胞在羰基应激条件下会发生一系列显著的生理生化改变;而生长环境相对较恶劣的酵母却对羰基应激不敏感。
[Abstract]:Aging is the result of both genetic and environmental factors. According to the theory of carbonyl stress senescence, in the process of lipid peroxidation and non-enzymatic glycosylation induced by oxygen free radicals, a common intermediate product with reactive activity, active carbonyl compounds, is produced. They can be crosslinked with almost all biological macromolecules in organism. This process is the core process of aging in organism and even the biochemical essence of aging change in organism. In this dissertation, two extreme normal cells (red blood cells with weak viability in vitro and yeast living in vitro for a long time) were used as experimental models to explore their responses to carbonyl stress. One is that the viscosity of blood increases gradually with the prolongation of storage time, but the specific biochemical mechanism is still unclear. In order to explore the mechanism of the increase of blood viscosity, the changes of TBARS level, carbonylation degree of erythrocyte membrane protein and the fluorescence intensity of protein during blood storage were investigated. The results showed that with the increase of blood viscosity, mercaptan level and protein fluorescence intensity decreased gradually, while TBARS level and carbonylation degree of membrane protein increased gradually. These results suggest that active carbonyl compounds can easily attack the amino and mercapto groups of proteins. Carbonylation of erythrocyte membrane proteins induced by carbonyl stress may be an important mechanism of viscosity increase during blood storage. Secondly, in order to explore the response of yeast cells to carbonyl stress, yeast cells were treated with different concentrations of MDA solution. After overnight cultivation, the light density of yeast was measured by UV spectrophotometer at 600 nm. At the same time, the activity of yeast cells treated with MDA was determined by PI fluorescence staining (Ex530/Em590). The results showed that the growth and activity of yeast cells were not significantly inhibited even under high concentration of MDA. This indicates that yeast cells have a strong ability to resist carbonyl stress, but the specific mechanism of this resistance needs to be further studied. To sum up, red blood cells in the blood will undergo a series of significant physiological and biochemical changes under carbonyl stress, while yeast, which has a relatively bad growth environment, is not sensitive to carbonyl stress.
【学位授予单位】：湖南师范大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R363
本文编号：2227291
[Abstract]:Aging is the result of both genetic and environmental factors. According to the theory of carbonyl stress senescence, in the process of lipid peroxidation and non-enzymatic glycosylation induced by oxygen free radicals, a common intermediate product with reactive activity, active carbonyl compounds, is produced. They can be crosslinked with almost all biological macromolecules in organism. This process is the core process of aging in organism and even the biochemical essence of aging change in organism. In this dissertation, two extreme normal cells (red blood cells with weak viability in vitro and yeast living in vitro for a long time) were used as experimental models to explore their responses to carbonyl stress. One is that the viscosity of blood increases gradually with the prolongation of storage time, but the specific biochemical mechanism is still unclear. In order to explore the mechanism of the increase of blood viscosity, the changes of TBARS level, carbonylation degree of erythrocyte membrane protein and the fluorescence intensity of protein during blood storage were investigated. The results showed that with the increase of blood viscosity, mercaptan level and protein fluorescence intensity decreased gradually, while TBARS level and carbonylation degree of membrane protein increased gradually. These results suggest that active carbonyl compounds can easily attack the amino and mercapto groups of proteins. Carbonylation of erythrocyte membrane proteins induced by carbonyl stress may be an important mechanism of viscosity increase during blood storage. Secondly, in order to explore the response of yeast cells to carbonyl stress, yeast cells were treated with different concentrations of MDA solution. After overnight cultivation, the light density of yeast was measured by UV spectrophotometer at 600 nm. At the same time, the activity of yeast cells treated with MDA was determined by PI fluorescence staining (Ex530/Em590). The results showed that the growth and activity of yeast cells were not significantly inhibited even under high concentration of MDA. This indicates that yeast cells have a strong ability to resist carbonyl stress, but the specific mechanism of this resistance needs to be further studied. To sum up, red blood cells in the blood will undergo a series of significant physiological and biochemical changes under carbonyl stress, while yeast, which has a relatively bad growth environment, is not sensitive to carbonyl stress.
【学位授予单位】：湖南师范大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R363
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